Antiviral phosphinate compounds

ABSTRACT

The invention is related to anti-viral phosphinate compounds, compositions containing such compounds, and therapeutic methods that include the administration of such compounds, as well as to processes and intermediates useful for preparing such compounds.

PRIORITY OF INVENTION

This application is a divisional of pending U.S. application Ser. No.11/825,606 filed 6 Jul. 2007 which claims priority to U.S. ProvisionalApplication No. 60/832,908 filed 24 Jul. 2006 and to U.S. ProvisionalApplication No. 60/819,488 filed 7 Jul. 2006, all of which areincorporated by reference in their entireties for all purposes.

FIELD OF THE INVENTION

The invention relates generally to phosphinate compounds with HCVinhibitory activity.

BACKGROUND OF THE INVENTION

Hepatitis C is recognized as a chronic viral disease of the liver whichis characterized by liver disease. Although drugs targeting the liverare in wide use and have shown effectiveness, toxicity and other sideeffects have limited their usefulness. Inhibitors of HCV are useful tolimit the establishment and progression of infection by HCV as well asin diagnostic assays for HCV.

There is a need for new HCV therapeutic agents.

SUMMARY OF THE INVENTION

In one embodiment the invention provides a compound of the inventionwhich is a compound of formula I:

or a pharmaceutically acceptable salt, or prodrug thereof, wherein:

-   -   R¹ is independently selected from H, alkyl, alkenyl, alkynyl,        aryl, cycloalkyl, heterocycle, halogen, haloalkyl,        alkylsulfonamido, arylsulfonamido, —C(O)NHS(O)₂—, or —S(O)₂—,        optionally substituted with one or more A³;    -   R² is selected from,    -   a) —C(Y¹)(A³),    -   b) (C2-10)alkyl, (C3-7)cycloalkyl or        (C1-4)alkyl-(C3-7)cycloalkyl, where said cycloalkyl and        alkyl-cycloalkyl may be optionally mono-, di- or tri-substituted        with (C1-3)alkyl, or        -   where said alkyl, cycloalkyl and alkyl-cycloalkyl may            optionally be mono- or di-substituted with substituents            selected from hydroxy and O—(C1-4)alkyl, or        -   where each of said alkyl-groups may optionally be mono-, di-            or tri-substituted with halogen, or        -   where each of said cycloalkyl groups being 5-, 6- or            7-membered, one or two —CH₂-groups not being directly linked            to each other may be optionally substituted replaced by —O—            such that the O-atom is linked to the N atom to which R² is            attached via at least two C-atoms,    -   c) phenyl, (C1-3)alkyl-phenyl, heteroaryl or        (C1-3)alkyl-heteroaryl, wherein the heteroaryl-groups are 5- or        6-membered having from 1 to 3 heteroatoms selected from N, O and        S, wherein said phenyl and heteroaryl groups may optionally be        mono-, di- or trisubstituted with substituents selected from        halogen, —OH, (C1-4)alkyl, O—(C1-4)alkyl, S—(C1-4)alkyl, —NH₂,        —CF₃, —NH((C1-4)alkyl) and —N((C1-4)alkyl)₂, —CONH₂ and        —CONH—(C1-4)alkyl; and wherein said (C1-3)alkyl may optionally        be substituted with one or more halogen; or    -   d) —S(O)₂(A³);    -   R³H or (C1-6)alkyl;    -   Y¹ is independently O, S, N(A³), N(O)(A³), N(OA³), N(O)(OA³) or        N(N(A³)(A³));    -   Z is O, S, or NR³;    -   Z¹ is selected from the following structures:

-   -   R_(a) is H or (C1-6)alkoxy;    -   R_(b) is H, F, Cl, Br, I, or (C1-6)alkyl;    -   R_(c) is H, cyano, F, Cl, Br, I, —C(═O)NR_(d)R_(e),        (C1-6)alkoxy, or phenyl that is optionally substituted with one        or more F, Cl, Br, I, (C1-6)alkyl, or (C1-6)alkoxy;    -   R_(d) and R_(e) are each independently H or (C1-6)alkyl;    -   each L is independently CH or N;    -   Z^(2a) is H, (C1-10)alkyl, (C2-10)alkenyl, (C2-10)alkynyl,        wherein any carbon atom may optionally be replaced with a        heteroatom selected from O, S or N, or Z^(2a) optionally forms a        heterocycle with one or more R¹, R², Q¹, or A³;    -   Z^(2b) is H, (C1-6)alkyl, (C2-8)alkenyl, (C2-8)alkynyl;    -   Q¹ is (C1-8)alkyl, (C2-8)alkenyl, or (C2-8)alkynyl; or Q¹ and        Z^(2a) taken together with the atoms to which they are attached        form a heterocycle, which heterocycle may optionally be        substituted with one or more oxo (═O) or A³;    -   A³ is independently selected from PRT, H, —OH, —C(O)OH, cyano,        alkyl, alkenyl, alkynyl, amino, amido, imido, imino, halogen,        CF₃, CH₂CF₃, cycloalkyl, nitro, aryl, aralkyl, alkoxy, aryloxy,        heterocycle, —C(A²)₃, —C(A²)₂-C(O)A², —C(O)A², —C(O)OA², —O(A²),        —N(A²)₂, —S(A²), —CH₂P(Y¹)(A²)(OA²), —CH₂P(Y¹)(A²)(N(A²)₂),        —CH₂P(Y¹)(OA²)(OA²), —OCH₂P(Y¹)(OA²)(OA²), —OCH₂P(Y¹)(A²)(OA²),        —OCH₂P(Y¹)(A²)(N(A²)₂), —C(O)OCH₂P(Y¹)(OA²)(OA²),        —C(O)OCH₂P(Y¹)(A²)(OA²), —C(O)OCH₂P(Y¹)(A²)(N(A²)₂),        —CH₂P(Y¹)(OA²)(N(A²)₂), —OCH₂P(Y¹)(OA²)(N(A²)₂),        —C(O)OCH₂P(Y¹)(OA²)(N(A²)₂), —CH₂P(Y¹)(N(A²)₂)(N(A²)₂),        —C(O)OCH₂P(Y¹)(N(A²)₂)(N(A²)₂), —OCH₂P(Y¹)(N(A²)₂)(N(A²)₂),        —(CH₂)_(m)-heterocycle, —(CH₂)_(m)C(O)Oalkyl,        —O—(CH₂)_(m)—O—C(O)—Oalkyl, —O—(CH₂)_(r)—O—C(O)—(CH₂)_(m)-alkyl,        —(CH₂)O—C(O)—O-alkyl, —(CH₂)_(m)O—C(O)—O-cycloalkyl,        —N(H)C(Me)C(O)O-alkyl, or alkoxy arylsulfonamide,        -   wherein each A³ may be optionally substituted with 1 to 4        -   —R¹, —P(Y¹)(OA²)(OA²), —P(Y¹)(OA²)(N(A²)₂), —P(Y¹)(A²)(OA²),            —P(Y)(A²)(N(A²)₂), or P(Y¹)(N(A²)₂)(N(A²)₂), —C(═O)N(A²)₂),            halogen, alkyl, alkenyl, alkynyl, aryl, carbocycle,            heterocycle, aralkyl, aryl sulfonamide, aryl            alkylsulfonamide, aryloxy sulfonamide, aryloxy            alkylsulfonamide, aryloxy arylsulfonamide, alkyl            sulfonamide, alkyloxy sulfonamide, alkyloxy            alkylsulfonamide, arylthio, —(CH₂)_(m)heterocycle,            —(CH₂)_(m)—C(O)O-alkyl, —O(CH₂)_(m)OC(O)Oalkyl,            —O—(CH₂)_(m), —O—C(O)—(CH₂)_(m)-alkyl,            —(CH₂)_(m)—O—C(O)—O-alkyl, —(CH₂)_(m)—O—C(O)—O-cycloalkyl,            —N(H)C(CH₃)C(O)O-alkyl, or alkoxy arylsulfonamide,            optionally substituted with R¹;    -   Optionally each independent instance of A³ and Q¹ can be taken        together with one or more A³ or Q¹ groups to form a ring;    -   A² is independently selected from PRT, H, alkyl, alkenyl,        alkynyl, amino, amino acid, alkoxy, aryloxy, cyano, haloalkyl,        cycloalkyl, aryl, heteroaryl, alkylsulfonamide, or        arylsulfonamide, optionally substituted with A³; and    -   m is 0 to 6.

The present invention also provides a pharmaceutical compositioncomprising a compound of the invention and at least one pharmaceuticallyacceptable carrier.

The present invention also provides a pharmaceutical composition for usein treating disorders associated with HCV.

The present invention also provides a pharmaceutical composition furthercomprising a nucleoside analog.

The present invention also provides for a pharmaceutical compositionfurther comprising an interferon or pegylated interferon.

The present invention also provides for a pharmaceutical compositionwherein said nucleoside analogue is selected from ribavirin, viramidinelevovirin, a L-nucleoside, and isatoribine and said interferon isa-interferon or pegylated interferon.

The present invention also provides for a method of treating disordersassociated with hepatitis C, said method comprising administering to anindividual a pharmaceutical composition which comprises atherapeutically effective amount of a compound of the invention.

The present invention also provides a method of inhibiting HCV,comprising administering to a mammal afflicted with a conditionassociated with HCV activity, an amount of a compound of the invention,effective to inhibit HCV.

The present invention also provides a compound of the invention for usein medical therapy (preferably for use in inhibiting HCV or treating acondition associated with HCV activity), as well as the use of acompound of the invention for the manufacture of a medicament useful forinhibiting HCV or the treatment of a condition associated with HCVactivity in a mammal.

The present invention also provides synthetic processes and novelintermediates disclosed herein which are useful for preparing compoundsof the invention. Some of the compounds of the invention are useful toprepare other compounds of the invention.

In another aspect the invention provides a method of inhibiting HCVactivity in a sample comprising treating the sample with a compound ofthe invention.

In one embodiment the invention provides a compound having improvedinhibitory or pharmacokinetic properties, including enhanced activityagainst development of viral resistance, improved oral bioavailability,greater potency or extended effective half-life in vivo. Certaincompounds of the invention may have fewer side effects, less complicateddosing schedules, or be orally active.

DETAILED DESCRIPTION OF THE INVENTION

Reference will now be made in detail to certain embodiments of theinvention, examples of which are illustrated in the accompanyingstructures and formulas. While the invention will be described inconjunction with the enumerated embodiments, it will be understood thatthey are not intended to limit the invention to those embodiments. Onthe contrary, the invention is intended to cover all alternatives,modifications, and equivalents, which may be included within the scopeof the present invention as defined by the embodiments.

Compounds of the Invention

The compounds of the invention exclude compounds heretofore known.However it is within the invention to use compounds that previously werenot known to have antiviral properties for antiviral purposes (e.g. toproduce an anti-viral effect in an animal). With respect to the UnitedStates, the compounds or compositions herein exclude compounds that areanticipated under 35 USC §102 or that are obvious under 35 USC §103.

Whenever a compound described herein is substituted with more than oneof the same designated group, e.g., “R¹” or “A³”, then it will beunderstood that the groups may be the same or different, i.e., eachgroup is independently selected.

“Alkyl” is C₁-C₁₈ hydrocarbon containing normal, secondary, tertiary orcyclic carbon atoms. Examples are methyl (Me, —CH₃), ethyl (Et,—CH₂CH₃), 1-propyl (n-Pr, n-propyl, —CH₂CH₂CH₃), 2-propyl (i-Pr,i-propyl, —CH(CH₃)₂), 1-butyl (n-Bu, n-butyl, —CH₂CH₂CH₂CH₃),2-methyl-1-propyl (i-Bu, i-butyl, —CH₂CH(CH₃)₂), 2-butyl (s-Bu, s-butyl,—CH(CH₃)CH₂CH₃), 2-methyl-2-propyl (t-Bu, t-butyl, —C(CH₃)₃), 1-pentyl(n-pentyl, —CH₂CH₂CH₂CH₂CH₃), 2-pentyl (—CH(CH₃)CH₂CH₂CH 3), 3-pentyl(—CH(CH₂CH₃)₂), 2-methyl-2-butyl (—C(CH₃)₂CH₂CH₃), 3-methyl-2-butyl(—CH(CH₃)CH(CH₃)₂), 3-methyl-1-butyl (—CH₂CH₂CH(CH₃)₂), 2-methyl-1-butyl(—CH₂CH(CH₃)CH₂CH₃), 1-hexyl (—CH₂CH₂CH₂CH₂CH₂CH₃)₂), 2-hexyl(—CH(CH₃)CH₂CH₂CH₂CH₃), 3-hexyl (—CH(CH₂CH₃)(CH₂CH₂CH₃)),2-methyl-2-pentyl (—C(CH₃)₂CH₂CH₂CH₃), 3-methyl-2-pentyl(—CH(CH₃)CH(CH₃)CH₂CH₃), 4-methyl-2-pentyl (—CH(CH₃)CH₂CH(CH₃)₂),3-methyl-3-pentyl (—C(CH₃)(CH₂CH₃)₂), 2-methyl-3-pentyl(—CH(CH₂CH₃)CH(CH₃)₂), 2,3-dimethyl-2-butyl (—C(CH₃)₂CH(CH₃)₂),3,3-dimethyl-2-butyl (—CH(CH₃)C(CH₃)₃.

“Alkenyl” is C₂-C₁₈ hydrocarbon containing normal, secondary, tertiaryor cyclic carbon atoms with at least one site of unsaturation, i.e. acarbon-carbon, sp² double bond. Examples include, but are not limitedto, ethylene or vinyl (—CH═CH₂), allyl (—CH₂CH═CH₂), cyclopentenyl(—C₅H₇), and 5-hexenyl (—CH₂ CH₂CH₂CH₂CH═CH₂).

“Alkynyl” is C₂-C₁₈ hydrocarbon containing normal, secondary, tertiaryor cyclic carbon atoms with at least one site of unsaturation, i.e. acarbon-carbon, sp triple bond. Examples include, but are not limited to,acetylenic (—C≡CH) and propargyl (—CH₂C≡CH).

“Alkylene” refers to a saturated, branched or straight chain or cyclichydrocarbon radical of 1-18 carbon atoms, and having two monovalentradical centers derived by the removal of two hydrogen atoms from thesame or two different carbon atoms of a parent alkane. Typical alkyleneradicals include, but are not limited to, methylene (—CH₂—) 1,2-ethyl(—CH₂CH₂—), 1,3-propyl (—CH₂CH₂CH₂—), 1,4-butyl (—CH₂CH₂CH₂CH₂—), andthe like.

“Alkenylene” refers to an unsaturated, branched or straight chain orcyclic hydrocarbon radical of 2-18 carbon atoms, and having twomonovalent radical centers derived by the removal of two hydrogen atomsfrom the same or two different carbon atoms of a parent alkene. Typicalalkenylene radicals include, but are not limited to, 1,2-ethylene(—CH═CH—).

“Alkynylene” refers to an unsaturated, branched or straight chain orcyclic hydrocarbon radical of 2-18 carbon atoms, and having twomonovalent radical centers derived by the removal of two hydrogen atomsfrom the same or two different carbon atoms of a parent alkyne. Typicalalkynylene radicals include, but are not limited to, acetylene (—C≡C—),propargyl (—CH₂C≡C—), and 4-pentynyl (—CH₂CH₂CH₂C≡CH—).

“Aryl” means a monovalent aromatic hydrocarbon radical of 6-20 carbonatoms derived by the removal of one hydrogen atom from a single carbonatom of a parent aromatic ring system. Typical aryl groups include, butare not limited to, radicals derived from benzene, substituted benzene,naphthalene, anthracene, biphenyl, and the like.

“Arylalkyl” refers to an acyclic alkyl radical in which one of thehydrogen atoms bonded to a carbon atom, typically a terminal or sp³carbon atom, is replaced with an aryl radical. Typical arylalkyl groupsinclude, but are not limited to, benzyl, 2-phenylethan-1-yl,naphthylmethyl, 2-naphthylethan-1-yl, naphthobenzyl,2-naphthophenylethan-1-yl and the like. The arylalkyl group comprises 6to 20 carbon atoms, e.g., the alkyl moiety, including alkanyl, alkenylor alkynyl groups, of the arylalkyl group is 1 to 6 carbon atoms and thearyl moiety is 5 to 14 carbon atoms.

“Substituted alkyl”, “substituted aryl”, and “substituted arylalkyl”mean alkyl, aryl, and arylalkyl respectively, in which one or morehydrogen atoms are each independently replaced with a non-hydrogensubstituent. Typical substituents include, but are not limited to, —X,—R, —O⁻, —OR, —SR, —S⁻, —NR₂, —NR₃, ═NR, —CX₃, —CN, —OCN, —SCN, —N═C═O,—NCS, —NO, —NO₂, ═N₂, —N₃, NC(═O)R, —C(═O)R, —C(═O)NRR—S(═O)₂O⁻,—S(═O)₂OH, —S(═O)₂R, —OS(═O)₂OR, —S(═O)₂NR, —S(═O)R, —OP(═O)O₂RR,—P(═O)O₂RR—P(═O)(O⁻)₂, —P(═O)(OH)₂, —C(═O)R, —C(═O)X, —C(S)R, —C(O)OR,—C(O)O⁻, —C(S)OR, —C(O)SR, —C(S)SR, —C(O)NRR, —C(S)NRR, —C(NR)NRR, whereeach X is independently a halogen: F, Cl, Br, or I; and each R isindependently —H, alkyl, aryl, heterocycle, protecting group or prodrugmoiety. Alkylene, alkenylene, and alkynylene groups may also besimilarly substituted.

“Heterocycle” as used herein includes by way of example and notlimitation these heterocycles described in Paquette, Leo A.; Principlesof Modern Heterocyclic Chemistry (W.A. Benjamin, New York, 1968),particularly Chapters 1, 3, 4, 6, 7, and 9; The Chemistry ofHeterocyclic Compounds, A Series of Monographs” (John Wiley & Sons, NewYork, 1950 to present), in particular Volumes 13, 14, 16, 19, and 28;and J. Am. Chem. Soc. (1960) 82:5566. In one specific embodiment of theinvention “heterocycle” includes a “carbocycle” as defined herein,wherein one or more (e.g. 1, 2, 3, or 4) carbon atoms have been replacedwith a heteroatom (e.g. O, N, or S).

Examples of heterocycles include by way of example and not limitationpyridyl, dihydroypyridyl, tetrahydropyridyl (piperidyl), thiazolyl,tetrahydrothiophenyl, sulfur oxidized tetrahydrothiophenyl, pyrimidinyl,furanyl, thienyl, pyrrolyl, pyrazolyl, imidazolyl, tetrazolyl,benzofuranyl, thianaphthalenyl, indolyl, indolenyl, quinolinyl,isoquinolinyl, benzimidazolyl, piperidinyl, 4-piperidonyl, pyrrolidinyl,2-pyrrolidonyl, pyrrolinyl, tetrahydrofuranyl, tetrahydroquinolinyl,tetrahydroisoquinolinyl, decahydroquinolinyl, octahydroisoquinolinyl,azocinyl, triazinyl, 6H-1,2,5-thiadiazinyl, 2H,6H-1,5,2-dithiazinyl,thienyl, thianthrenyl, pyranyl, isobenzofuranyl, chromenyl, xanthenyl,phenoxathinyl, 2H-pyrrolyl, isothiazolyl, isoxazolyl, pyrazinyl,pyridazinyl, indolizinyl, isoindolyl, 3H-indolyl, 1H-indazoly, purinyl,4H-quinolizinyl, phthalazinyl, naphthyridinyl, quinoxalinyl,quinazolinyl, cinnolinyl, pteridinyl, 4H-carbazolyl, carbazolyl,β-carbolinyl, phenanthridinyl, acridinyl, pyrimidinyl, phenanthrolinyl,phenazinyl, phenothiazinyl, furazanyl, phenoxazinyl, isochromanyl,chromanyl, imidazolidinyl, imidazolinyl, pyrazolidinyl, pyrazolinyl,piperazinyl, indolinyl, isoindolinyl, quinuclidinyl, morpholinyl,oxazolidinyl, benzotriazolyl, benzisoxazolyl, oxindolyl, benzoxazolinyl,isatinoyl, and bis-tetrahydrofuranyl:

By way of example and not limitation, carbon bonded heterocycles arebonded at position 2, 3, 4, 5, or 6 of a pyridine, position 3, 4, 5, or6 of a pyridazine, position 2, 4, 5, or 6 of a pyrimidine, position 2,3, 5, or 6 of a pyrazine, position 2, 3, 4, or 5 of a furan,tetrahydrofuran, thiofuran, thiophene, pyrrole or tetrahydropyrrole,position 2, 4, or 5 of an oxazole, imidazole or thiazole, position 3, 4,or 5 of an isoxazole, pyrazole, or isothiazole, position 2 or 3 of anaziridine, position 2, 3, or 4 of an azetidine, position 2, 3, 4, 5, 6,7, or 8 of a quinoline or position 1, 3, 4, 5, 6, 7, or 8 of anisoquinoline. Still more typically, carbon bonded heterocycles include2-pyridyl, 3-pyridyl, 4-pyridyl, 5-pyridyl, 6-pyridyl, 3-pyridazinyl,4-pyridazinyl, 5-pyridazinyl, 6-pyridazinyl, 2-pyrimidinyl,4-pyrimidinyl, 5-pyrimidinyl, 6-pyrimidinyl, 2-pyrazinyl, 3-pyrazinyl,5-pyrazinyl, 6-pyrazinyl, 2-thiazolyl, 4-thiazolyl, or 5-thiazolyl.

By way of example and not limitation, nitrogen bonded heterocycles arebonded at position 1 of an aziridine, azetidine, pyrrole, pyrrolidine,2-pyrroline, 3-pyrroline, imidazole, imidazolidine, 2-imidazoline,3-imidazoline, pyrazole, pyrazoline, 2-pyrazoline, 3-pyrazoline,piperidine, piperazine, indole, indoline, 1H-indazole, position 2 of aisoindole, or isoindoline, position 4 of a morpholine, and position 9 ofa carbazole, or β-carboline. Still more typically, nitrogen bondedheterocycles include 1-aziridyl, 1-azetedyl, 1-pyrrolyl, 1-imidazolyl,1-pyrazolyl, and 1-piperidinyl.

“Carbocycle” refers to a saturated, unsaturated or aromatic ring havingup to about 25 carbon atoms. Typically, a carbocycle has about 3 to 7carbon atoms as a monocycle, about 7 to 12 carbon atoms as a bicycle,and up to about 25 carbon atoms as a polycycle. Monocyclic carbocyclestypically have 3 to 6 ring atoms, still more typically 5 or 6 ringatoms. Bicyclic carbocycles typically have 7 to 12 ring atoms, e.g.,arranged as a bicyclo [4,5], [5,5], [5,6] or [6,6] system, or 9 or 10ring atoms arranged as a bicyclo [5,6] or [6,6] system. The termcarbocycle includes “cycloalkyl” which is a saturated or unsaturatedcarbocycle. Examples of monocyclic carbocycles include cyclopropyl,cyclobutyl, cyclopentyl, 1-cyclopent-1-enyl, 1-cyclopent-2-enyl,1-cyclopent-3-enyl, cyclohexyl, 1-cyclohex-1-enyl, 1-cyclohex-2-enyl,1-cyclohex-3-enyl, phenyl, spiryl and naphthyl. When Q¹ and Z^(2a) takentogether with the atoms to which they are attached form a heterocycle,the heterocycle formed by Q¹ and Z^(2a) taken together with the atoms towhich they are attached may typically comprise up to about 25 atoms.

The term “chiral” refers to molecules which have the property ofnon-superimposability of the mirror image partner, while the term“achiral” refers to molecules which are superimposable on their mirrorimage partner.

The term “stereoisomers” refers to compounds which have identicalchemical constitution, but differ with regard to the arrangement of theatoms or groups in space.

“Diastereomer” refers to a stereoisomer with two or more centers ofchirality and whose molecules are not mirror images of one another.Diastereomers have different physical properties, e.g., melting points,boiling points, spectral properties, and reactivities. Mixtures ofdiastereomers may separate under high resolution analytical proceduressuch as electrophoresis and chromatography.

“Enantiomers” refer to two stereoisomers of a compound which arenon-superimposable mirror images of one another.

The term “treatment” or “treating,” to the extent it relates to adisease or condition includes preventing the disease or condition fromoccurring, inhibiting the disease or condition, eliminating the diseaseor condition, and/or relieving one or more symptoms of the disease orcondition.

The term “PRT” is selected from the terms “prodrug moiety” and“protecting group” as defined herein.

Stereochemical definitions and conventions used herein generally followS. P. Parker, Ed., McGraw-Hill Dictionary of Chemical Terms (1984)McGraw-Hill Book Company, New York; and Eliel, E. and Wilen, S.,Stereochemistry of Organic Compounds (1994) John Wiley & Sons, Inc., NewYork. Many organic compounds exist in optically active forms, i.e., theyhave the ability to rotate the plane of plane-polarized light. Indescribing an optically active compound, the prefixes D and L or R and Sare used to denote the absolute configuration of the molecule about itschiral center(s). The prefixes d and l or (+) and (−) are employed todesignate the sign of rotation of plane-polarized light by the compound,with (−) or 1 meaning that the compound is levorotatory. A compoundprefixed with (+) or d is dextrorotatory. For a given chemicalstructure, these stereoisomers are identical except that they are mirrorimages of one another. A specific stereoisomer may also be referred toas an enantiomer, and a mixture of such isomers is often called anenantiomeric mixture. A 50:50 mixture of enantiomers is referred to as aracemic mixture or a racemate, which may occur where there has been nostereoselection or stereospecificity in a chemical reaction or process.The terms “racemic mixture” and “racemate” refer to an equimolar mixtureof two enantiomeric species, devoid of optical activity. The inventionincludes all stereoisomers of the compounds described herein.

Prodrugs

The term “prodrug” as used herein refers to any compound that whenadministered to a biological system generates the drug substance, i.e.active ingredient, as a result of spontaneous chemical reaction(s),enzyme catalyzed chemical reaction(s), photolysis, and/or metabolicchemical reaction(s). A prodrug is thus a covalently modified analog orlatent form of a therapeutically-active compound.

“Prodrug moiety” refers to a labile functional group which separatesfrom the active inhibitory compound during metabolism, systemically,inside a cell, by hydrolysis, enzymatic cleavage, or by some otherprocess (Bundgaard, Hans, “Design and Application of Prodrugs” in ATextbook of Drug Design and Development (1991), P. Krogsgaard-Larsen andH. Bundgaard, Eds. Harwood Academic Publishers, pp. 113-191). Enzymeswhich are capable of an enzymatic activation mechanism with thephosphonate prodrug compounds of the invention include, but are notlimited to, amidases, esterases, microbial enzymes, phospholipases,cholinesterases, and phosphases. Prodrug moieties can serve to enhancesolubility, absorption and lipophilicity to optimize drug delivery,bioavailability and efficacy. A prodrug moiety may include an activemetabolite or drug itself.

Exemplary prodrug moieties include the hydrolytically sensitive orlabile acyloxymethyl esters —CH₂OC(═O)R⁹ and acyloxymethyl carbonates—CH₂OC(═O)OR⁹ where R⁹ is C₁-C₆ alkyl, C₁-C₆ substituted alkyl, C₆-C₂₀aryl or C₆-C₂₀ substituted aryl. The acyloxyalkyl ester was first usedas a prodrug strategy for carboxylic acids and then applied tophosphates and phosphonates by Farquhar et al. (1983) J. Pharm. Sci. 72:324; also U.S. Pat. Nos. 4,816,570, 4,968,788, 5,663,159 and 5,792,756.Subsequently, the acyloxyalkyl ester was used to deliver phosphonicacids across cell membranes and to enhance oral bioavailability. A closevariant of the acyloxyalkyl ester, the alkoxycarbonyloxyalkyl ester(carbonate), may also enhance oral bioavailability as a prodrug moietyin the compounds of the combinations of the invention. An exemplaryacyloxymethyl ester is pivaloyloxymethoxy, (POM) —CH₂C(═O)C(CH₃)₃. Anexemplary acyloxymethyl carbonate prodrug moiety ispivaloyloxymethylcarbonate (POC)—CH₂C(═O)OC(CH₃)₃.

Aryl esters of phosphorus groups, especially phenyl esters, are reportedto enhance oral bioavailability (De Lombaert et al. (1994) J. Med. Chem.37: 498). Phenyl esters containing a carboxylic ester ortho to aphosphate have also been described (Khamnei and Torrence, (1996) J. Med.Chem. 39:4109-4115). Benzyl esters are reported to generate parentphosphonic acids. In some cases, substituents at the ortho- orpara-position may accelerate the hydrolysis. Benzyl analogs with anacylated phenol or an alkylated phenol may generate the phenoliccompound through the action of enzymes, e.g., esterases, oxidases, etc.,which in turn undergoes cleavage at the benzylic C—O bond to generatephosphoric acid and a quinone methide intermediate. Examples of thisclass of prodrugs are described by Mitchell et al. (1992) J. Chem. Soc.Perkin Trans. II 2345; Glazier WO 91/19721. Still other benzylicprodrugs have been described containing a carboxylic ester-containinggroup attached to the benzylic methylene (Glazier WO 91/19721).Thio-containing prodrugs are reported to be useful for the intracellulardelivery of phosphonate drugs. These proesters contain an ethylthiogroup in which the thiol group is either esterified with an acyl groupor combined with another thiol group to form a disulfide.Deesterification or reduction of the disulfide generates the free thiointermediate which subsequently breaks down to the phosphoric acid andepisulfide (Puech et al. (1993) Antiviral Res., 22: 155-174; Benzaria etal. (1996) J. Med. Chem. 39: 4958).

Protecting Groups

In the context of the present invention, protecting groups includeprodrug moieties and chemical protecting groups.

“Protecting group” refers to a moiety of a compound that masks or altersthe properties of a functional group or the properties of the compoundas a whole. Chemical protecting groups and strategies forprotection/deprotection are well known in the art. See e.g., ProtectiveGroups in Organic Chemistry, Theodora W. Greene, John Wiley & Sons,Inc., New York, 1991. Protecting groups are often utilized to mask thereactivity of certain functional groups, to assist in the efficiency ofdesired chemical reactions, e.g., making and breaking chemical bonds inan ordered and planned fashion. Protection of functional groups of acompound alters other physical properties besides the reactivity of theprotected functional group, such as the polarity, lipophilicity(hydrophobicity), and other properties which can be measured by commonanalytical tools. Chemically protected intermediates may themselves bebiologically active or inactive.

Protected compounds may also exhibit altered, and in some cases,optimized properties in vitro and in vivo, such as passage throughcellular membranes and resistance to enzymatic degradation orsequestration. In this role, protected compounds with intendedtherapeutic effects may be referred to as prodrugs. Another function ofa protecting group is to convert the parental drug into a prodrug,whereby the parental drug is released upon conversion of the prodrug invivo. Because active prodrugs may be absorbed more effectively than theparental drug, prodrugs may possess greater potency in vivo than theparental drug. Protecting groups are removed either in vitro, in theinstance of chemical intermediates, or in vivo, in the case of prodrugs.With chemical intermediates, it is not particularly important that theresulting products after deprotection, e.g., alcohols, bephysiologically acceptable, although in general it is more desirable ifthe products are pharmacologically innocuous.

Protecting groups are available, commonly known and used, and areoptionally used to prevent side reactions with the protected groupduring synthetic procedures, i.e. routes or methods to prepare thecompounds of the invention. For the most part the decision as to whichgroups to protect, when to do so, and the nature of the chemicalprotecting group “PG” will be dependent upon the chemistry of thereaction to be protected against (e.g., acidic, basic, oxidative,reductive or other conditions) and the intended direction of thesynthesis. The PG groups do not need to be, and generally are not, thesame if the compound is substituted with multiple PG. In general, PGwill be used to protect functional groups such as carboxyl, hydroxyl,thio, or amino groups and to thus prevent side reactions or to otherwisefacilitate the synthetic efficiency. The order of deprotection to yieldfree, deprotected groups is dependent upon the intended direction of thesynthesis and the reaction conditions to be encountered, and may occurin any order as determined by the artisan.

Various functional groups of the compounds of the invention may beprotected. For example, protecting groups for —OH groups (whetherhydroxyl, carboxylic acid, phosphonic acid, or other functions) include“ether- or ester-forming groups”. Ether- or ester-forming groups arecapable of functioning as chemical protecting groups in the syntheticschemes set forth herein. However, some hydroxyl and thio protectinggroups are neither ether- nor ester-forming groups, as will beunderstood by those skilled in the art, and are included with amides,discussed below.

A very large number of hydroxyl protecting groups and amide-forminggroups and corresponding chemical cleavage reactions are described inProtective Groups in Organic Synthesis, Theodora W. Greene (John Wiley &Sons, Inc., New York, 1991, ISBN 0-471-62301-6) (“Greene”). See alsoKocienski, Philip J.; Protecting Groups (Georg Thieme Verlag Stuttgart,New York, 1994), which is incorporated by reference in its entiretyherein. In particular Chapter 1, Protecting Groups: An Overview, pages1-20, Chapter 2, Hydroxyl Protecting Groups, pages 21-94, Chapter 3,Diol Protecting Groups, pages 95-117, Chapter 4, Carboxyl ProtectingGroups, pages 118-154, Chapter 5, Carbonyl Protecting Groups, pages155-184. For protecting groups for carboxylic acid, phosphonic acid,phosphonate, sulfonic acid and other protecting groups for acids seeGreene as set forth below.

A³ and A² may be H, alkyl, or an ether- or ester-forming group.“Ether-forming group” means a group which is capable of forming astable, covalent bond between the parental molecule and a group havingthe formula:

Wherein V_(a) is a tetravalent atom typically selected from C and Si;V_(b) is a trivalent atom typically selected from B, Al, N, and P, moretypically N and P; V_(c) is a divalent atom typically selected from O,S, and Se, more typically S; V₁ is a group bonded to V_(a), V_(b) orV_(c) by a stable, single covalent bond, typically V₁ is A² groups; V₂is a group bonded to V_(a) or V_(b) by a stable, double covalent bond,provided that V₂ is not ═O, ═S or ═N—, typically V₂ is ═C(V₁)₂ whereinV₁ is as described above; and V₃ is a group bonded to V_(a) by a stable,triple covalent bond, typically V₃ is ∫C(V₁) wherein V₁ is as describedabove.

“Ester-forming group” means a group which is capable of forming astable, covalent bond between the parental molecule and a group havingthe formula:

Wherein V_(a), V_(b), and V₁, are as described above; V_(d) is apentavalent atom typically selected from P and N; V_(e) is a hexavalentatom typically S; and V₄ is a group bonded to V_(a), V_(b), V_(d) orV_(e) by a stable, double covalent bond, provided that at least one V₄is ═O, ═S or ═N—V₁, typically V₄, when other than ═O, ═S or ═N—, is═C(V₁)₂ wherein V₁ is as described above.

Protecting groups for —OH functions (whether hydroxy, acid or otherfunctions) are embodiments of “ether- or ester-forming groups”.Particularly of interest are ether- or ester-forming groups that arecapable of functioning as protecting groups in the synthetic schemes setforth herein. However, some hydroxyl and thio protecting groups areneither ether- nor ester-forming groups, as will be understood by thoseskilled in the art, and are included with amides, discussed below, andare capable of protecting hydroxyl or thio groups such that hydrolysisfrom the parental molecule yields hydroxyl or thio.

In its ester-forming role, A³ or A² typically is bound to any acidicgroup such as, by way of example and not limitation, a —CO₂H or —C(S)OHgroup, thereby resulting in —CO₂A² or —CO₂A³. A² for example is deducedfrom the enumerated ester groups of WO 95/07920.

Examples of A² include

C₃-C₁₂ heterocycle (described above) or aryl. These aromatic groupsoptionally are polycyclic or monocyclic. Examples include phenyl,spiryl, 2- and 3-pyrrolyl, 2- and 3-thienyl, 2- and 4-imidazolyl, 2-, 4-and 5-oxazolyl, 3- and 4-isoxazolyl, 2-, 4- and 5-thiazolyl, 3-, 4- and5-isothiazolyl, 3- and 4-pyrazolyl, 1-, 2-, 3- and 4-pyridinyl, and 1-,2-, 4- and 5-pyrimidinyl, C₃-C₁₂ heterocycle or aryl substituted withhalo, R¹, R₁—O—C₁-C₁₂ alkylene, C₁-C₁₂ alkoxy, CN, NO₂, OH, carboxy,carboxyester, thiol, thioester, C₁-C₁₂ haloalkyl (1-6 halogen atoms),C₂-C₁₂ alkenyl or C₂-C₁₂ alkynyl. Such groups include 2-, 3- and4-alkoxyphenyl (C₁-C₁₂ alkyl), 2-, 3- and 4-methoxyphenyl, 2-, 3- and4-ethoxyphenyl, 2,3-, 2,4-, 2,5-, 2,6-, 3,4- and 3,5-diethoxyphenyl, 2-and 3-carboethoxy-4-hydroxyphenyl, 2- and 3-ethoxy-4-hydroxyphenyl, 2-and 3-ethoxy-5-hydroxyphenyl, 2- and 3-ethoxy-6-hydroxyphenyl, 2-,3- and4-O-acetylphenyl, 2-, 3- and 4-dimethylaminophenyl, 2-, 3- and4-methylmercaptophenyl, 2-, 3- and 4-halophenyl (including 2-, 3- and4-fluorophenyl and 2-, 3- and 4-chlorophenyl), 2,3-, 2,4-, 2,5-, 2,6-,3,4- and 3,5-dimethylphenyl, 2,3-, 2,4-, 2,5-, 2,6-, 3,4- and3,5-biscarboxyethylphenyl, 2,3-, 2,4-, 2,5-, 2,6-, 3,4- and3,5-dimethoxyphenyl, 2,3-, 2,4-, 2,5-, 2,6-, 3,4- and 3,5-dihalophenyl(including 2,4-difluorophenyl and 3,5-difluorophenyl), 2-, 3- and4-haloalkylphenyl (1 to 5 halogen atoms, C₁-C₁₂ alkyl including4-trifluoromethylphenyl), 2-, 3- and 4-cyanophenyl, 2-, 3- and4-nitrophenyl, 2-, 3- and 4-haloalkylbenzyl (1 to 5 halogen atoms,C₁-C₁₂ alkyl including 4-trifluoromethylbenzyl and 2-, 3- and4-trichloromethylphenyl and 2-, 3- and 4-trichloromethylphenyl),4-N-methylpiperidinyl, 3-N-methylpiperidinyl, 1-ethylpiperazinyl,benzyl, alkylsalicylphenyl (C₁-C₄ alkyl, including 2-, 3- and4-ethylsalicylphenyl), 2-,3- and 4-acetylphenyl, 1,8-dihydroxynaphthyl(—C₁₀H₆—OH) and aryloxy ethyl [C₆-C₉ aryl (including phenoxy ethyl)],2,2′-dihydroxybiphenyl, 2-, 3- and 4-N,N-dialkylaminophenol,—C₆H₄—CH₂—N(CH₃)₂, trimethoxybenzyl, triethoxybenzyl, 2-alkyl pyridinyl(C1-4 alkyl);

C₄-C₈ esters of 2-carboxyphenyl; and C₁-C₄ alkylene-C₃-C₆ aryl(including benzyl, —CH₂-pyrrolyl, —CH₂-thienyl, —CH₂-imidazolyl,—CH₂-oxazolyl, —CH₂-isoxazolyl, —CH₂-thiazolyl, —CH₂-isothiazolyl,—CH₂-pyrazolyl, —CH₂-pyridinyl and —CH₂-pyrimidinyl) substituted in thearyl moiety by 3 to 5 halogen atoms or 1 to 2 atoms or groups selectedfrom halogen, C₁-C₁₂ alkoxy (including methoxy and ethoxy), cyano,nitro, OH, C₁-C₁₂ haloalkyl (1 to 6 halogen atoms; including —CH₂—CCl₃),C₁-C₁₂ alkyl (including methyl and ethyl), C₂-C₁₂ alkenyl or C₂-C₁₂alkynyl;

alkoxy ethyl [C₁-C₆ alkyl including —CH₂—CH₂—O—CH₃ (methoxy ethyl)];

alkyl substituted by any of the groups set forth above for aryl, inparticular OH or by 1 to 3 halo atoms (including —CH₃, —CH(CH₃)₂,

—C(CH₃)₃, —CH₂CH₃, —(CH₂)₂CH₃, —(CH₂)₃CH₃, —(CH₂)₄CH₃, —(CH₂)₅CH₃,—CH₂CH₂F, —CH₂CH₂Cl, —CH₂CF₃, and —CH₂CCl₃);

—N-2-propylmorpholino, 2,3-dihydro-6-hydroxyindene, sesamol, catecholmonoester, —CH₂—C(O)—N(R¹)₂, —CH₂—S(O)(R¹), —CH₂—S(O)²(R¹),—CH₂—CH(OC(O)CH₂R¹)—CH₂(OC(O)CH₂R¹), cholesteryl, enolpyruvate(HOOC—C(═CH₂)—), glycerol;

a 5 or 6 carbon monosaccharide, disaccharide or oligosaccharide (3 to 9monosaccharide residues);

triglycerides such as a-D-β-diglycerides (wherein the fatty acidscomposing glyceride lipids generally are naturally occurring saturatedor unsaturated C₆₋₂₆, C₆₋₁₈ or C₆₋₁₀ fatty acids such as linoleic,lauric, myristic, palmitic, stearic, oleic, palmitoleic, linolenic andthe like fatty acids) linked to acyl of the parental compounds hereinthrough a glyceryl oxygen of the triglyceride;

phospholipids linked to the carboxyl group through the phosphate of thephospholipid;

phthalidyl (shown in FIG. 1 of Clayton et al., Antimicrob. Agents Chemo.5(6):670-671 [1974]);

cyclic carbonates such as (5-R_(d)-2-oxo-1,3-dioxolen-4-yl)methyl esters(Sakamoto et al., Chem. Pharm. Bull. 32(6)2241-2248 [1984]) where R_(d)is R₁, R₄ or aryl; and

The hydroxyl groups of the compounds of this invention optionally aresubstituted with one of groups III, IV or V disclosed in WO94/21604, orwith isopropyl.

As further embodiments, Table A lists examples of A² ester moieties thatfor example can be bonded via oxygen to —C(O)O— and —P(O)(R)(O—) groups.Several amidates also are shown, which are bound directly to —C(O)— or—P(O)₂. Esters of structures 1-5, 8-10 and 16, 17, 19-22 are synthesizedby reacting the compound herein having a free hydroxyl with thecorresponding halide (chloride or acyl chloride and the like) andN,N-dicyclohexyl-N-morpholine carboxamidine (or another base such asDBU, triethylamine, CsCO₃, N,N-dimethylaniline and the like) in DMF (orother solvent such as acetonitrile or N-methylpyrrolidone). When A³ isphosphonate, the esters of structures 5-7, 11, 12, 21, and 23-26 aresynthesized by reaction of the alcohol or alkoxide salt (or thecorresponding amines in the case of compounds such as 13, 14 and 15)with the monochlorophosphonate or dichlorophosphonate (or anotheractivated phosphonate).

TABLE A  1. —CH₂—C(O)—N(R₁)₂ *  2. —CH₂—S(O)(R₁)  3. —CH₂—S(O)₂(R₁)  4.—CH₂—O—C(O)—CH₂—C₆H₅  5. 3-cholesteryl  6. 3-pyridyl  7.N-ethylmorpholino  8. —CH₂—O—C(O)—C₆H₅  9. —CH₂—O—C(O)—CH₂CH₃ 10.—CH₂—O—C(O)—C(CH₃)₃ 11. —CH₂—CCl₃ 12. —C₆H₅ 13. —NH—CH₂—C(O)O—CH₂CH₃ 14.—N(CH₃)—CH₂—C(O)O—CH₂CH₃ 15. —NHR₁ 16. —CH₂—O—C(O)—C₁₀H₁₅ 17.—CH₂—O—C(O)—CH(CH₃)₂ 18. —CH₂—C#H(OC(O)CH₂R₁)—CH₂—(OC(O)CH₂R₁)* 19.

20.

21.

22.

23.

24.

25.

26.

#-chiral center is (R), (S) or racemate.

Other esters that are suitable for use herein are described in EuropeanPatent No. 632,048.

A² also includes “double ester” forming profunctionalities such as—CH₂OC(O)OCH₃,

—CH₂SCOCH₃, —CH₂OCON(CH₃)₂, or alkyl- or aryl-acyloxyalkyl groups of thestructure —CH(R¹)O((CO)R₃₇) or —CH(R¹)((CO)OR₃₈) (linked to oxygen ofthe acidic group) wherein R₃₇ and R₃₈ are alkyl, aryl, or alkylarylgroups (see U.S. Pat. No. 4,968,788). Frequently R₃₇ and R₃₈ are bulkygroups such as branched alkyl, ortho-substituted aryl, meta-substitutedaryl, or combinations thereof, including normal, secondary, iso- andtertiary alkyls of 1-6 carbon atoms. An example is the pivaloyloxymethylgroup. These are of particular use with prodrugs for oraladministration. Examples of such useful A² groups are alkylacyloxymethylesters and their derivatives, including —CH(CH₂CH₂OCH₃)OC(O)C(CH₃)₃,

—CH₂OC(O)C₁₀H₁₅, —CH₂OC(O)C(CH₃)₃, —CH(CH₂OCH₃)OC(O)C(CH₃)₃,—CH(CH(CH₃)₂)OC(O)C(CH₃)₃, —CH₂OC(O)CH₂CH(CH₃)₂, —CH₂OC(O)C₆H₁₁,—CH₂OC(O)C₆H₅, —CH₂OC(O)C₁₀H₁₅, —CH₂OC(O)CH₂CH₃, —CH₂OC(O)CH(CH₃)₂,—CH₂OC(O)C(CH₃)₃ and —CH₂OC(O)CH₂C₆H₅.

For prodrug purposes, the ester typically chosen is one heretofore usedfor antiviral drugs, in particular the cyclic carbonates, double esters,or the phthalidyl, aryl or alkyl esters.

As noted A³ or A² groups optionally are used to prevent side reactionswith the protected group during synthetic procedures, so they functionas protecting groups (PRT) during synthesis. For the most part thedecision as to which groups to protect, when to do so, and the nature ofthe PRT will be dependent upon the chemistry of the reaction to beprotected against (e.g., acidic, basic, oxidative, reductive or otherconditions) and the intended direction of the synthesis. The PRT groupsdo not need to be, and generally are not, the same if the compound issubstituted with multiple PRT. In general, PRT will be used to protectcarboxyl, hydroxyl or amino groups. The order of deprotection to yieldfree groups is dependent upon the intended direction of the synthesisand the reaction conditions to be encountered, and may occur in anyorder as determined by the artisan.

In some embodiments the A² protected acidic group is an ester of theacidic group and A² is the residue of a hydroxyl-containingfunctionality. In other embodiments, an amino compound is used toprotect the acid functionality. The residues of suitable hydroxyl oramino-containing functionalities are set forth above or are found in WO95/07920. Of particular interest are the residues of amino acids, aminoacid esters, polypeptides, or aryl alcohols. Typical amino acid,polypeptide and carboxyl-esterified amino acid residues are described onpages 11-18 and related text of WO 95/07920 as groups L1 or L2. WO95/07920 expressly teaches the amidates of phosphonic acids, but it willbe understood that such amidates are formed with any of the acid groupsset forth herein and the amino acid residues set forth in WO 95/07920.

Typical A² esters for protecting A³ acidic functionalities are alsodescribed in WO 95/07920, again understanding that the same esters canbe formed with the acidic groups herein as with the phosphonate of the'920 publication. Typical ester groups are defined at least on WO95/07920 pages 89-93 (under R³¹ or R³⁵), the table on page 105, andpages 21-23 (as R¹). Of particular interest are esters of unsubstitutedaryl such as phenyl or arylalkyl such benzyl, or hydroxy-, halo-,alkoxy-, carboxy- and/or alkylestercarboxy-substituted aryl oralkylaryl, especially phenyl, ortho-ethoxyphenyl, or C₁-C₄alkylestercarboxyphenyl (salicylate C₁-C₁₂ alkylesters).

The protected acidic groups A³, particularly when using the esters oramides of WO 95/07920, are useful as prodrugs for oral administration.However, it is not essential that the A³ acidic group be protected inorder for the compounds of this invention to be effectively administeredby the oral route. When the compounds of the invention having protectedgroups, in particular amino acid amidates or substituted andunsubstituted aryl esters are administered systemically or orally theyare capable of hydrolytic cleavage in vivo to yield the free acid.

One or more of the acidic hydroxyls are protected. If more than oneacidic hydroxyl is protected then the same or a different protectinggroup is employed, e.g., the esters may be different or the same, or amixed amidate and ester may be used.

Typical A² hydroxy protecting groups described in Greene (pages 14-118)include Ethers (Methyl); Substituted Methyl Ethers (Methoxymethyl,Methylthiomethyl, t-Butylthiomethyl, (Phenyldimethylsilyl)methoxymethyl,Benzyloxymethyl, p-Methoxybenzyloxymethyl, (4-Methoxyphenoxy)methyl,Guaiacolmethyl, t-Butoxymethyl, 4-Pentenyloxymethyl, Siloxymethyl,2-Methoxyethoxymethyl, 2,2,2-Trichloroethoxymethyl,Bis(2-chloroethoxy)methyl, 2-(Trimethylsilyl)ethoxymethyl,Tetrahydropyranyl, 3-Bromotetrahydropyranyl, Tetrahydropthiopyranyl,1-Methoxycyclohexyl, 4-Methoxytetrahydropyranyl,4-Methoxytetrahydrothiopyranyl, 4-MethoxytetrahydropthiopyranylS,S-Dioxido, 1-[(2-Chloro-4-methyl)phenyl]-4-methoxypiperidin-4-yl,35,1,4-Dioxan-2-yl, Tetrahydrofuranyl, Tetrahydrothiofuranyl,2,3,3a,4,5,6,7,7a-Octahydro-7,8,8-trimethyl-4,7-methanobenzofuran-2-yl));Substituted Ethyl Ethers (1-Ethoxyethyl, 1-(2-Chloroethoxy)ethyl,1-Methyl-1-methoxyethyl, 1-Methyl-1-benzyloxyethyl,1-Methyl-1-benzyloxy-2-fluoroethyl, 2,2,2-Trichloroethyl,2-Trimethylsilylethyl, 2-(Phenylselenyl)ethyl, t-Butyl, Allyl,p-Chlorophenyl, p-Methoxyphenyl, 2,4-Dinitrophenyl, Benzyl); SubstitutedBenzyl Ethers (p-Methoxybenzyl, 3,4-Dimethoxybenzyl, o-Nitrobenzyl,p-Nitrobenzyl, p-Halobenzyl, 2,6-Dichlorobenzyl, p-Cyanobenzyl,p-Phenylbenzyl, 2- and 4-Picolyl, 3-Methyl-2-picolyl N-Oxido,Diphenylmethyl, p,p′-Dinitrobenzhydryl, 5-Dibenzosuberyl,Triphenylmethyl, a-Naphthyldiphenylmethyl,p-methoxyphenyldiphenylmethyl, Di(p-methoxyphenyl)phenylmethyl,Tri(p-methoxyphenyl)methyl, 4-(4′-Bromophenacyloxy)phenyldiphenylmethyl,4,4′,4″-Tris(4,5-dichlorophthalimidophenyl)methyl,4,4′,4″-Tris(levulinoyloxyphenyl)methyl,4,4′,4″-Tris(benzoyloxyphenyl)methyl,3-(Imidazol-1-ylmethyl)bis(4′,4″-dimethoxyphenyl)methyl,1,1-Bis(4-methoxyphenyl)-1′-pyrenylmethyl, 9-Anthryl,9-(9-Phenyl)xanthenyl, 9-(9-Phenyl-10-oxo)anthryl,1,3-Benzodithiolan-2-yl, Benzisothiazolyl S,S-Dioxido); Silyl Ethers(Trimethylsilyl, Triethylsilyl, Triisopropylsilyl,Dimethylisopropylsilyl, Diethylisopropylsilyl, Dimethylthexylsilyl,t-Butyldimethylsilyl, t-Butyldiphenylsilyl, Tribenzylsilyl,Tri-p-xylylsilyl, Triphenylsilyl, Diphenylmethylsilyl,t-Butylmethoxyphenylsilyl); Esters (Formate, Benzoylformate, Acetate,Chloroacetate, Dichloroacetate, Trichloroacetate, Trifluoroacetate,Methoxyacetate, Triphenylmethoxyacetate, Phenoxyacetate,p-Chlorophenoxyacetate, p-poly-Phenylacetate, 3-Phenylpropionate,4-Oxopentanoate (Levulinate), 4,4-(Ethylenedithio)pentanoate, Pivaloate,Adamantoate, Crotonate, 4-Methoxycrotonate, Benzoate, p-Phenylbenzoate,2,4,6-Trimethylbenzoate (Mesitoate)); Carbonates (Methyl,9-Fluorenylmethyl, Ethyl, 2,2,2-Trichloroethyl, 2-(Trimethylsilyl)ethyl,2-(Phenylsulfonyl)ethyl, 2-(Triphenylphosphonio)ethyl, Isobutyl, Vinyl,Allyl, p-Nitrophenyl, Benzyl, p-Methoxybenzyl, 3,4-Dimethoxybenzyl,o-Nitrobenzyl, p-Nitrobenzyl, S-Benzyl Thiocarbonate,4-Ethoxy-1-naphthyl, Methyl Dithiocarbonate); Groups With AssistedCleavage (2-Iodobenzoate, 4-Azidobutyrate, 4-Nitro-4-methylpentanoate,o-(Dibromomethyl)benzoate, 2-Formylbenzenesulfonate,2-(Methylthiomethoxy)ethyl Carbonate, 4-(Methylthiomethoxy)butyrate,2-(Methylthiomethoxymethyl)benzoate); Miscellaneous Esters(2,6-Dichloro-4-methylphenoxyacetate, 2,6-Dichloro-4-(1,1,3,3tetramethylbutyl)phenoxyacetate,2,4-Bis(1,1-dimethylpropyl)phenoxyacetate, Chlorodiphenylacetate,Isobutyrate, Monosuccinate, (E)-2-Methyl-2-butenoate (Tigloate),o-(Methoxycarbonyl)benzoate, p-poly-Benzoate, a-Naphthoate, Nitrate,Alkyl N,N,N′,N′-Tetramethylphosphorodiamidate, N-Phenylcarbamate,Borate, Dimethylphosphinothioyl, 2,4-Dinitrophenylsulfenate); andSulfonates (Sulfate, Methanesulfonate (Mesylate), Benzylsulfonate,Tosylate).

More typically, A² hydroxy protecting groups include substituted methylethers, substituted benzyl ethers, silyl ethers, and esters includingsulfonic acid esters, still more typically, trialkylsilyl ethers,tosylates and acetates.

Typical 1,2-diol protecting groups (thus, generally where two OH groupsare taken together with the A² protecting functionality) are describedin Greene at pages 118-142 and include Cyclic Acetals and Ketals(Methylene, Ethylidene, 1-t-Butylethylidene, 1-Phenylethylidene,(4-Methoxyphenyl)ethylidene, 2,2,2-Trichloroethylidene, Acetonide(Isopropylidene), Cyclopentylidene, Cyclohexylidene, Cycloheptylidene,Benzylidene, p-Methoxybenzylidene, 2,4-Dimethoxybenzylidene,3,4-Dimethoxybenzylidene, 2-Nitrobenzylidene); Cyclic Ortho Esters(Methoxymethylene, Ethoxymethylene, Dimethoxymethylene,1-Methoxyethylidene, 1-Ethoxyethylidine, 1,2-Dimethoxyethylidene,a-Methoxybenzylidene, 1-(N,N-Dimethylamino)ethylidene Derivative,a-(N,N-Dimethylamino)benzylidene Derivative, 2-Oxacyclopentylidene);Silyl Derivatives (Di-t-butylsilylene Group,1,3-(1,1,3,3-Tetraisopropyldisiloxanylidene), andTetra-t-butoxydisiloxane-1,3-diylidene), Cyclic Carbonates, CyclicBoronates, Ethyl Boronate and Phenyl Boronate.

More typically, 1,2-diol protecting groups include those shown in TableB, still more typically, epoxides, acetonides, cyclic ketals and arylacetals.

TABLE B

wherein R⁹ is C₁-C₆ alkyl.

A² is also H, a protecting group for amino or the residue of acarboxyl-containing compound, in particular H, —C(O)R₄, an amino acid, apolypeptide or a protecting group not —C(O)R₄, amino acid orpolypeptide. Amide-forming A² are found for instance in group A³. WhenA² is an amino acid or polypeptide it has the structureR₁₅NHCH(R₁₆)C(O)—, where R₁₅ is H, an amino acid or polypeptide residue,or R₁₅, and R₁₆ is defined below.

R₁₆ is lower alkyl or lower alkyl (C₁-C₆) substituted with amino,carboxyl, amide, carboxyl ester, hydroxyl, C₆-C₇ aryl, guanidinyl,imidazolyl, indolyl, sulfhydryl, sulfoxide, and/or alkylphosphate. R₁₀also is taken together with the amino acid aN to form a proline residue(R₁₀═(CH₂)₃—). However, R₁₀ is generally the side group of anaturally-occurring amino acid such as H, —CH₃, —CH(CH₃)₂,—CH₂—CH(CH₃)₂, —CHCH₃—CH₂—CH₃, —CH₂—C₆H₅, —CH₂CH₂—S—CH₃, —CH₂OH,—CH(OH)—CH₃, —CH₂—SH, —CH₂—C₆H₄OH, —CH₂—CO—NH₂, —CH₂—CH₂—CO—NH₂,—CH₂—COOH, —CH₂—CH₂—COOH, —(CH₂)₄—NH₂ and —(CH₂)₃—NH—C(NH₂)—NH₂. R₁₀also includes 1-guanidinoprop-3-yl, benzyl, 4-hydroxybenzyl,imidazol-4-yl, indol-3-yl, methoxyphenyl and ethoxyphenyl. A² areresidues of carboxylic acids for the most part, but any of the typicalamino protecting groups described by Greene at pages 315-385 are useful.They include Carbamates (methyl and ethyl, 9-fluorenylmethyl,9-(2-sulfo)fluoroenylmethyl, 9-(2,7-dibromo)fluorenylmethyl,2,7-di-t-buthyl-[9-(10,10-dioxo-10,10,10,10-tetrahydrothioxanthyl)]methyl,4-methoxyphenacyl); Substituted Ethyl (2,2,2-trichloroethyl,2-trimethylsilylethyl, 2-phenylethyl, 1-(1-adamantyl)-1-methylethyl,1,1-dimethyl-2-haloethyl, 1,1-dimethyl-2,2-dibromoethyl,1,1-dimethyl-2,2,2-trichloroethyl, 1-methyl-1-(4-biphenylyl)ethyl,1-(3,5-di-t-butylphenyl)-1-methylethyl, 2-(2′- and 4′-pyridyl)ethyl,2-(N,N-dicyclohexylcarboxamido)ethyl, t-butyl, 1-adamantyl, vinyl,allyl, 1-isopropylallyl, cinnamyl, 4-nitrocinnamyl, 8-quinolyl,N-hydroxypiperidinyl, alkyldithio, benzyl, p-methoxybenzyl,p-nitrobenzyl, p-bromobenzyl, p-chlorobenzyl, 2,4-dichlorobenzyl,4-methylsulfinylbenzyl, 9-anthrylmethyl, diphenylmethyl); Groups WithAssisted Cleavage (2-methylthioethyl, 2-methylsulfonylethyl,2-(p-toluenesulfonyl)ethyl, [2-(1,3-dithianyl)]methyl,4-methylthiophenyl, 2,4-dimethylthiophenyl, 2-phosphonioethyl,2-triphenylphosphonioisopropyl, 1,1-dimethyl-2-cyanoethyl,m-choro-p-acyloxybenzyl, p-(dihydroxyboryl)benzyl,5-benzisoxazolylmethyl, 2-(trifluoromethyl)-6-chromonylmethyl); GroupsCapable of Photolytic Cleavage (m-nitrophenyl, 3,5-dimethoxybenzyl,o-nitrobenzyl, 3,4-dimethoxy-6-nitrobenzyl,phenyl(o-nitrophenyl)methyl); Urea-Type Derivatives(phenothiazinyl-(10)-carbonyl, N′-p-toluenesulfonylaminocarbonyl,N′-phenylaminothiocarbonyl); Miscellaneous Carbamates (t-amyl, S-benzylthiocarbamate, p-cyanobenzyl, cyclobutyl, cyclohexyl, cyclopentyl,cyclopropylmethyl, p-decyloxybenzyl, diisopropylmethyl,2,2-dimethoxycarbonylvinyl, o-(N,N-dimethylcarboxamido)benzyl,1,1-dimethyl-3-(N,N-dimethylcarboxamido)propyl, 1,1-dimethylpropynyl,di(2-pyridyl)methyl, 2-furanylmethyl, 2-Iodoethyl, Isobornyl, Isobutyl,Isonicotinyl, p-(p′-Methoxyphenylazo)benzyl, 1-methylcyclobutyl,1-methylcyclohexyl, 1-methyl-1-cyclopropylmethyl,1-methyl-1-(3,5-dimethoxyphenyl)ethyl,1-methyl-1-(p-phenylazophenyl)ethyl, 1-methyl-1-phenylethyl,1-methyl-1-(4-pyridyl)ethyl, phenyl, p-(phenylazo)benzyl,2,4,6-tri-t-butylphenyl, 4-(trimethylammonium)benzyl,2,4,6-trimethylbenzyl); Amides (N-formyl, N-acetyl, N-choroacetyl,N-trichoroacetyl, N-trifluoroacetyl, N-phenylacetyl,N-3-phenylpropionyl, N-picolinoyl, N-3-pyridylcarboxamide,N-benzoylphenylalanyl, N-benzoyl, N-p-phenylbenzoyl); Amides WithAssisted Cleavage (N-o-nitrophenylacetyl, N-o-nitrophenoxyacetyl,N-acetoacetyl, (N′-dithiobenzyloxycarbonylamino)acetyl,N-3-(p-hydroxyphenyl)propionyl, N-3-(o-nitrophenyl)propionyl,N-2-methyl-2-(o-nitrophenoxy)propionyl,N-2-methyl-2-(o-phenylazophenoxy)propionyl, N-4-chlorobutyryl,N-3-methyl-3-nitrobutyryl, N-o-nitrocinnamoyl, N-acetylmethionine,N-o-nitrobenzoyl, N-o-(benzoyloxymethyl)benzoyl,4,5-diphenyl-3-oxazolin-2-one); Cyclic Imide Derivatives (N-phthalimide,N-dithiasuccinoyl, N-2,3-diphenylmaleoyl, N-2,5-dimethylpyrrolyl,N-1,1,4,4-tetramethyldisilylazacyclopentane adduct, 5-substituted1,3-dimethyl-1,3,5-triazacyclohexan-2-one, 5-substituted1,3-dibenzyl-1,3-5-triazacyclohexan-2-one, 1-substituted3,5-dinitro-4-pyridonyl); N-Alkyl and N-Aryl Amines (N-methyl, N-allyl,N-[2-(trimethylsilyl)ethoxy]methyl, N-3-acetoxypropyl,N-(1-isopropyl-4-nitro-2-oxo-3-pyrrolin-3-yl), Quaternary AmmoniumSalts, N-benzyl, N-di(4-methoxyphenyl)methyl, N-5-dibenzosuberyl,N-triphenylmethyl, N-(4-methoxyphenyl)diphenylmethyl,N-9-phenylfluorenyl, N-2,7-dichloro-9-fluorenylmethylene,N-ferrocenylmethyl, N-2-picolylamine N′-oxide), Imine Derivatives(N-1,1-dimethylthiomethylene, N-benzylidene, N-p-methoxybenzylidene,N-diphenylmethylene, N-[(2-pyridyl)mesityl]methylene,N,(N′,N′-dimethylaminomethylene, N,N′-isopropylidene,N-p-nitrobenzylidene, N-salicylidene, N-5-chlorosalicylidene,N-(5-chloro-2-hydroxyphenyl)phenylmethylene, N-cyclohexylidene); EnamineDerivatives (N-(5,5-dimethyl-3-oxo-1-cyclohexenyl)); N-Metal Derivatives(N-borane derivatives, N-diphenylborinic acid derivatives,N-[phenyl(pentacarbonylchromium- or -tungsten)]carbenyl, N-copper orN-zinc chelate); N—N Derivatives (N-nitro, N-nitroso, N-oxide); N—PDerivatives (N-diphenylphosphinyl, N-dimethylthiophosphinyl,N-diphenylthiophosphinyl, N-dialkyl phosphoryl, N-dibenzyl phosphoryl,N-diphenyl phosphoryl); N—Si Derivatives; N—S Derivatives; N-SulfenylDerivatives (N-benzenesulfenyl, N-o-nitrobenzenesulfenyl,N-2,4-dinitrobenzenesulfenyl, N-pentachlorobenzenesulfenyl,N-2-nitro-4-methoxybenzenesulfenyl, N-triphenylmethylsulfenyl,N-3-nitropyridinesulfenyl); and N-sulfonyl Derivatives(N-p-toluenesulfonyl, N-benzenesulfonyl,N-2,3,6-trimethyl-4-methoxybenzenesulfonyl,N-2,4,6-trimethoxybenzenesulfonyl,N-2,6-dimethyl-4-methoxybenzenesulfonyl, N-pentamethylbenzenesulfonyl,N-2,3,5,6-tetramethyl-4-methoxybenzenesulfonyl,N-4-methoxybenzenesulfonyl, N-2,4,6-trim ethylbenzenesulfonyl,N-2,6-dimethoxy-4-methylbenzenesulfonyl,N-2,2,5,7,8-pentamethylchroman-6-sulfonyl, N-methanesulfonyl,N-□-trimethylsilyethanesulfonyl, N-9-anthracenesulfonyl,N-4-(4′,8′-dimethoxynaphthylmethyl)benzenesulfonyl, N-benzylsulfonyl,N-trifluoromethylsulfonyl, N-phenacylsulfonyl).

More typically, protected amino groups include carbamates and amides,still more typically, —NHC(O)R¹ or —N═CR¹N(R¹)₂. Another protectinggroup, also useful as a prodrug at the A³ site, particularly for aminoor —NH(R₅), is:

see for example Alexander, J. et al.; J. Med. Chem. 1996, 39, 480-486.

A² is also H or the residue of an amino-containing compound, inparticular an amino acid, a polypeptide, a protecting group, —NHSO₂R₄,NHC(O)R₄, —N(R₄)₂, NH₂ or —NH(R₄)(H), whereby for example the carboxylor phosphonic acid groups of A³ are reacted with the amine to form anamide, as in —C(O)A², —P(O)(A²)₂ or —P(O)(OH)(A²). In general, A² hasthe structure R₁₇C(O)CH(R₁₆)NH—, where R₁₇ is OH, OA², OR₅, an aminoacid or a polypeptide residue.

Amino acids are low molecular weight compounds, on the order of lessthan about 1,000 MW, that contain at least one amino or imino group andat least one carboxyl group. Generally the amino acids will be found innature, i.e., can be detected in biological material such as bacteria orother microbes, plants, animals or man. Suitable amino acids typicallyare alpha amino acids, i.e. compounds characterized by one amino orimino nitrogen atom separated from the carbon atom of one carboxyl groupby a single substituted or unsubstituted alpha carbon atom. Ofparticular interest are hydrophobic residues such as mono- or di-alkylor aryl amino acids, cycloalkylamino acids and the like. These residuescontribute to cell permeability by increasing the partition coefficientof the parental drug. Typically, the residue does not contain asulfhydryl or guanidino substituent.

Naturally-occurring amino acid residues are those residues foundnaturally in plants, animals or microbes, especially proteins thereof.Polypeptides most typically will be substantially composed of suchnaturally-occurring amino acid residues. These amino acids are glycine,alanine, valine, leucine, isoleucine, serine, threonine, cysteine,methionine, glutamic acid, aspartic acid, lysine, hydroxylysine,arginine, histidine, phenylalanine, tyrosine, tryptophan, proline,asparagine, glutamine and hydroxyproline.

When A² are single amino acid residues or polypeptides they usually aresubstituted at A³. These conjugates are produced by forming an amidebond between a carboxyl group of the amino acid (or C-terminal aminoacid of a polypeptide for example) and amino nitrogen. Similarly,conjugates are formed between A³ and an amino group of an amino acid orpolypeptide. Generally, only one of any site in the parental molecule isamidated with an amino acid as described herein, although it is withinthe scope of this invention to introduce amino acids at more than onepermitted site. Usually, a carboxyl group of A³ is amidated with anamino acid. In general, the a-amino or a-carboxyl group of the aminoacid or the terminal amino or carboxyl group of a polypeptide are bondedto the parental functionalities, i.e., carboxyl or amino groups in theamino acid side chains generally are not used to form the amide bondswith the parental compound (although these groups may need to beprotected during synthesis of the conjugates as described furtherbelow).

With respect to the carboxyl-containing side chains of amino acids orpolypeptides it will be understood that the carboxyl group optionallywill be blocked, e.g. by A², esterified with A² or amidated with A².Similarly, the amino side chains R₁₆ optionally will be blocked with A²or substituted with R₅.

Such ester or amide bonds with side chain amino or carboxyl groups, likethe esters or amides with the parental molecule, optionally arehydrolyzable in vivo or in vitro under acidic (pH<3) or basic (pH>10)conditions. Alternatively, they are substantially stable in thegastrointestinal tract of humans but are hydrolyzed enzymatically inblood or in intracellular environments. The esters or amino acid orpolypeptide amidates also are useful as intermediates for thepreparation of the parental molecule containing free amino or carboxylgroups. The free acid or base of the parental compound, for example, isreadily formed from the esters or amino acid or polypeptide conjugatesof this invention by conventional hydrolysis procedures.

When an amino acid residue contains one or more chiral centers, any ofthe D, L, meso, threo or erythro (as appropriate) racemates, scalematesor mixtures thereof may be used. In general, if the intermediates are tobe hydrolyzed non-enzymatically (as would be the case where the amidesare used as chemical intermediates for the free acids or free amines), Disomers are useful. On the other hand, L isomers are more versatilesince they can be susceptible to both non-enzymatic and enzymatichydrolysis, and are more efficiently transported by amino acid ordipeptidyl transport systems in the gastrointestinal tract.

Examples of suitable amino acids whose residues are represented by A²include the following:

Glycine;

Aminopolycarboxylic acids, e.g., aspartic acid, β-hydroxyaspartic acid,glutamic acid, β-hydroxyglutamic acid, β-methylaspartic acid,β-methylglutamic acid, β,β-dimethylaspartic acid, γ-hydroxyglutamicacid, β,γ-dihydroxyglutamic acid, β-phenylglutamic acid,γ-methyleneglutamic acid, 3-aminoadipic acid, 2-aminopimelic acid,2-aminosuberic acid and 2-aminosebacic acid;

Amino acid amides such as glutamine and asparagine;

Polyamino- or polybasic-monocarboxylic acids such as arginine, lysine,β-aminoalanine, γ-aminobutyrine, ornithine, citruline, homoarginine,homocitrulline, hydroxylysine, allohydroxylsine and diaminobutyric acid;

Other basic amino acid residues such as histidine;

Diaminodicarboxylic acids such as a,a′-diaminosuccinic acid,a,a′-diaminoglutaric acid, a,a′-diaminoadipic acid, a,a′-diaminopimelicacid, a,a′-diamino-β-hydroxypimelic acid, a,a′-diaminosuberic acid,a,a′-diaminoazelaic acid, and a,a′-diaminosebacic acid;

Imino acids such as proline, hydroxyproline, allohydroxyproline,γ-methylproline, pipecolic acid, 5-hydroxypipecolic acid, andazetidine-2-carboxylic acid;

A mono- or di-alkyl (typically C₁-C₈ branched or normal) amino acid suchas alanine, valine, leucine, allylglycine, butyrine, norvaline,norleucine, heptyline, a-methylserine, a-amino-a-methyl-γ-hydroxyvalericacid, a-amino-a-methyl-d-hydroxyvaleric acid,a-amino-a-methyl-e-hydroxycaproic acid, isovaline, a-methylglutamicacid, a-aminoisobutyric acid, a-aminodiethylacetic acid,a-aminodiisopropylacetic acid, a-aminodi-n-propylacetic acid,a-aminodiisobutylacetic acid, a-aminodi-n-butylacetic acid,a-aminoethylisopropylacetic acid, a-amino-n-propylacetic acid,a-aminodiisoamyacetic acid, a-methylaspartic acid, a-methylglutamicacid, 1-aminocyclopropane-1-carboxylic acid, isoleucine, alloisoleucine,tert-leucine, β-methyltryptophan and a-amino-β-ethyl-β-phenylpropionicacid;

-   -   β-phenylserinyl;    -   Aliphatic a-amino-β-hydroxy acids such as serine,        β-hydroxyleucine, β-hydroxynorleucine, β-hydroxynorvaline, and        a-amino-β-hydroxystearic acid;    -   a-Amino, a-, γ-, d- or e-hydroxy acids such as homoserine,        γ-hydroxynorvaline, d-hydroxynorvaline and        epsilon-hydroxynorleucine residues; canavine and canaline;        γ-hydroxyornithine;    -   2-hexosaminic acids such as D-glucosaminic acid or        D-galactosaminic acid;    -   a-Amino-β-thiols such as penicillamine, β-thiolnorvaline or        β-thiolbutyrine;    -   Other sulfur containing amino acid residues including cysteine;        homocystine, β-phenylmethionine, methionine, S-allyl-L-cysteine        sulfoxide, 2-thiolhistidine, cystathionine, and thiol ethers of        cysteine or homocysteine;    -   Phenylalanine, tryptophan and ring-substituted a-amino acids        such as the phenyl- or cyclohexylamino acids a-aminophenylacetic        acid, a-aminocyclohexylacetic acid and        a-amino-β-cyclohexylpropionic acid; phenylalanine analogues and        derivatives comprising aryl, lower alkyl, hydroxy, guanidino,        oxyalkylether, nitro, sulfur or halo-substituted phenyl (e.g.,        tyrosine, methyltyrosine and o-chloro-, p-chloro-, 3,4-dichloro,        o-, m- or p-methyl-, 2,4,6-trimethyl-, 2-ethoxy-5-nitro-,        2-hydroxy-5-nitro- and p-nitro-phenylalanine); furyl-, thienyl-,        pyridyl-, pyrimidinyl-, purinyl- or naphthyl-alanines; and        tryptophan analogues and derivatives including kynurenine,        3-hydroxykynurenine, 2-hydroxytryptophan and        4-carboxytryptophan;    -   a-Amino substituted amino acids including sarcosine        (N-methylglycine), N-benzylglycine, N-methylalanine,        N-benzylalanine, N-methylphenylalanine, N-benzylphenylalanine,        N-methylvaline and N-benzylvaline; and    -   a-Hydroxy and substituted a-hydroxy amino acids including        serine, threonine, allothreonine, phosphoserine and        phosphothreonine.

Polypeptides are polymers of amino acids in which a carboxyl group ofone amino acid monomer is bonded to an amino or imino group of the nextamino acid monomer by an amide bond. Polypeptides include dipeptides,low molecular weight polypeptides (about 1500-5000 MW) and proteins.Proteins optionally contain 3, 5, 10, 50, 75, 100 or more residues, andsuitably are substantially sequence-homologous with human, animal, plantor microbial proteins. They include enzymes (e.g., hydrogen peroxidase)as well as immunogens such as KLH, or antibodies or proteins of any typeagainst which one wishes to raise an immune response. The nature andidentity of the polypeptide may vary widely.

The polypeptide amidates are useful as immunogens in raising antibodiesagainst either the polypeptide (if it is not immunogenic in the animalto which it is administered) or against the epitopes on the remainder ofthe compound of this invention.

Antibodies capable of binding to the parental non-peptidyl compound areused to separate the parental compound from mixtures, for example indiagnosis or manufacturing of the parental compound. The conjugates ofparental compound and polypeptide generally are more immunogenic thanthe polypeptides in closely homologous animals, and therefore make thepolypeptide more immunogenic for facilitating raising antibodies againstit. Accordingly, the polypeptide or protein may not need to beimmunogenic in an animal typically used to raise antibodies, e.g.,rabbit, mouse, horse, or rat, but the final product conjugate should beimmunogenic in at least one of such animals. The polypeptide optionallycontains a peptidolytic enzyme cleavage site at the peptide bond betweenthe first and second residues adjacent to the acidic heteroatom. Suchcleavage sites are flanked by enzymatic recognition structures, e.g. aparticular sequence of residues recognized by a peptidolytic enzyme.

Peptidolytic enzymes for cleaving the polypeptide conjugates of thisinvention are well known, and in particular include carboxypeptidases.Carboxypeptidases digest polypeptides by removing C-terminal residues,and are specific in many instances for particular C-terminal sequences.Such enzymes and their substrate requirements in general are well known.For example, a dipeptide (having a given pair of residues and a freecarboxyl terminus) is covalently bonded through its a-amino group to thephosphorus or carbon atoms of the compounds herein. In embodiments whereA³ is phosphonate it is expected that this peptide will be cleaved bythe appropriate peptidolytic enzyme, leaving the carboxyl of theproximal amino acid residue to autocatalytically cleave thephosphonoamidate bond.

Suitable dipeptidyl groups (designated by their single letter code) areAA, AR, AN, AD, AC, AE, AQ, AG, AH, AI, AL, AK, AM, AF, AP, AS, AT, AW,AY, AV, RA, RR, RN, RD, RC, RE, RQ, RG, RH, RI, RL, RK, RM, RF, RP, RS,RT, RW, RY, RV, NA, NR, NN, ND, NC, NE, NQ, NG, NH, NI, NL, NK, NM, NF,NP, NS, NT, NR, NY, NV, DA, DR, DN, DD, DC, DE, DQ, DG, DH, DI, DL, DK,DM, DF, DP, DS, DT, DW, DY, DV, CA, CR, CN, CD, CC, CE, CQ, CG, CH, CI,CL, CK, CM, CF, CP, CS, CT, CW, CY, CV, EA, ER, EN, ED, EC, EE, EQ, EG,EH, EI, EL, EK, EM, EF, EP, ES, ET, EW, EY, EV, QA, QR, QN, QD, QC, QE,QQ, QG, QH, QI, QL, QK, QM, QF, QP, QS, QT, QW, QY, QV, GA, GR, GN, GD,GC, GE, GQ, GG, GH, GI, GL, GK, GM, GF, GP, GS, GT, GW, GY, GV, HA, HR,HN, HD, HC, HE, HQ, HG, HH, HI, HL, HK, HM, HF, HP, HS, HT, HW, HY, HV,IA, IR, IN, ID, IC, IE, IQ, IG, IH, II, IL, IK, IM, IF, IP, IS, IT, IW,IY, IV, LA, LR, LN, LD, LC, LE, LQ, LG, LH, LI, LL, LK, LM, LF, LP, LS,LT, LW, LY, LV, KA, KR, KN, KD, KC, KE, KQ, KG, KH, KI, KL, KK, KM, KF,KP, KS, KT, KW, KY, KV, MA, MR, MN, MD, MC, ME, MQ, MG, MH, MI, ML, MK,MM, MF, MP, MS, MT, MW, MY, MV, FA, FR, FN, FD, FC, FE, FQ, FG, FH, FI,FL, FK, FM, FF, FP, FS, FT, FW, FY, FV, PA, PR, PN, PD, PC, PE, PQ, PG,PH, PI, PL, PK, PM, PF, PP, PS, PT, PW, PY, PV, SA, SR, SN, SD, SC, SE,SQ, SG, SH, SI, SL, SK, SM, SF, SP, SS, ST, SW, SY, SV, TA, TR, TN, TD,TC, TE, TQ, TG, TH, TI, TL, TK, TM, TF, TP, TS, TT, TW, TY, TV, WA, WR,WN, WD, WC, WE, WQ, WG, WH, WI, WL, WK, WM, WF, WP, WS, WT, WW, WY, WV,YA, YR, YN, YD, YC, YE, YQ, YG, YH, YI, YL, YK, YM, YF, YP, YS, YT, YW,YY, YV, VA, VR, VN, VD, VC, VE, VQ, VG, VH, VI, VL, VK, VM, VF, VP, VS,VT, VW, VY and VV.

Tripeptide residues are also useful as A². When A³ is phosphonate, thesequence —X⁴-pro-X⁵— (where X⁴ is any amino acid residue and X⁵ is anamino acid residue, a carboxyl ester of proline, or hydrogen) will becleaved by luminal carboxypeptidase to yield X⁴ with a free carboxyl,which in turn is expected to autocatalytically cleave thephosphonoamidate bond. The carboxy group of X⁵ optionally is esterifiedwith benzyl.

Dipeptide or tripeptide species can be selected on the basis of knowntransport properties and/or susceptibility to peptidases that can affecttransport to intestinal mucosal or other cell types. Dipeptides andtripeptides lacking an a-amino group are transport substrates for thepeptide transporter found in brush border membrane of intestinal mucosalcells (Bai, J. P. F., “Pharm Res.” 9:969-978 (1992). Transport competentpeptides can thus be used to enhance bioavailability of the amidatecompounds. Di- or tripeptides having one or more amino acids in the Dconfiguration are also compatible with peptide transport and can beutilized in the amidate compounds of this invention. Amino acids in theD configuration can be used to reduce the susceptibility of a di- ortripeptide to hydrolysis by proteases common to the brush border such asaminopeptidase N (EC 3.4.11.2). In addition, di- or tripeptidesalternatively are selected on the basis of their relative resistance tohydrolysis by proteases found in the lumen of the intestine. Forexample, tripeptides or polypeptides lacking asp and/or glu are poorsubstrates for aminopeptidase A (EC 3.4.11.7), di- or tripeptideslacking amino acid residues on the N-terminal side of hydrophobic aminoacids (leu, tyr, phe, val, trp) are poor substrates for endopeptidase24.11 (EC 3.4.24.11), and peptides lacking a pro residue at thepenultimate position at a free carboxyl terminus are poor substrates forcarboxypeptidase P (EC 3.4.17). Similar considerations can also beapplied to the selection of peptides that are either relativelyresistant or relatively susceptible to hydrolysis by cytosolic, renal,hepatic, serum or other peptidases. Such poorly cleaved polypeptideamidates are immunogens or are useful for bonding to proteins in orderto prepare immunogens.

HCV-Inhibitory Compounds

The compounds of the invention include those with HCV-inhibitoryactivity as well as intermediate compounds that are useful for preparingthe active compounds. The term “HCV-inhibitory compound” includes thosecompounds that inhibit HCV.

Typically, compounds of the invention have a molecular weight of fromabout 200 amu to about 10,000 amu; in a specific embodiment of theinvention, compounds have a molecular weight of less than about 5000amu; in another specific embodiment of the invention, compounds have amolecular weight of less than about 2500 amu; in another specificembodiment of the invention, compounds have a molecular weight of lessthan about 1000 amu; in another specific embodiment of the invention,compounds have a molecular weight of less than about 800 amu; in anotherspecific embodiment of the invention, compounds have a molecular weightof less than about 600 amu; and in another specific embodiment of theinvention, compounds have a molecular weight of less than about 600 amuand a molecular weight of greater than about 400 amu.

The compounds of the invention also typically have a log D(polarity)less than about 5. In one embodiment the invention provides compoundshaving a log D less than about 4; in another one embodiment theinvention provides compounds having a log D less than about 3; inanother one embodiment the invention provides compounds having a log Dgreater than about −5; in another one embodiment the invention providescompounds having a log D greater than about −3; and in another oneembodiment the invention provides compounds having a log D greater thanabout 0 and less than about 3.

Selected substituents within the compounds of the invention are presentto a recursive degree. In this context, “recursive substituent” meansthat a substituent may recite another instance of itself. Because of therecursive nature of such substituents, theoretically, a large number maybe present in any given embodiment. For example, R^(x) contains a R^(y)substituent. R^(y) can be R², which in turn can be R³. If R³ is selectedto be R^(3c), then a second instance of R^(x) can be selected. One ofordinary skill in the art of medicinal chemistry understands that thetotal number of such substituents is reasonably limited by the desiredproperties of the compound intended. Such properties include, by ofexample and not limitation, physical properties such as molecularweight, solubility or log P, application properties such as activityagainst the intended target, and practical properties such as ease ofsynthesis.

By way of example and not limitation, A³, A² and R¹ are all recursivesubstituents in certain embodiments. Typically, each of these mayindependently occur 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7,6, 5, 4, 3, 2, 1, or 0, times in a given embodiment. More typically,each of these may independently occur 12 or fewer times in a givenembodiment. Whenever a compound described herein is substituted withmore than one of the same designated group, e.g., “R¹” or “A³”, then itwill be understood that the groups may be the same or different, i.e.,each group is independently selected. Wavy lines indicate the site ofcovalent bond attachments to the adjoining groups, moieties, or atoms.

In one embodiment of the invention, the compound is in an isolated andpurified form. Generally, the term “isolated and purified” means thatthe compound is substantially free from biological materials (e.g.blood, tissue, cells, etc.). In one specific embodiment of theinvention, the term means that the compound or conjugate of theinvention is at least about 50 wt. % free from biological materials; inanother specific embodiment, the term means that the compound orconjugate of the invention is at least about 75 wt. % free frombiological materials; in another specific embodiment, the term meansthat the compound or conjugate of the invention is at least about 90 wt.% free from biological materials; in another specific embodiment, theterm means that the compound or conjugate of the invention is at leastabout 98 wt. % free from biological materials; and in anotherembodiment, the term means that the compound or conjugate of theinvention is at least about 99 wt. % free from biological materials. Inanother specific embodiment, the invention provides a compound orconjugate of the invention that has been synthetically prepared (e.g.,ex vivo).

Cellular Accumulation

In one embodiment, the invention provides compounds capable ofaccumulating in human PBMC (peripheral blood mononuclear cells). PBMCrefer to blood cells having round lymphocytes and monocytes.Physiologically, PBMC are critical components of the mechanism againstinfection. PBMC may be isolated from heparinized whole blood of normalhealthy donors or buffy coats, by standard density gradientcentrifugation and harvested from the interface, washed (e.g.phosphate-buffered saline) and stored in freezing medium. PBMC may becultured in multi-well plates. At various times of culture, supernatantmay be either removed for assessment, or cells may be harvested andanalyzed (Smith R. et al (2003) Blood 102(7):2532-2540). The compoundsof this embodiment may further comprise a phosphonate or phosphonateprodrug. More typically, the phosphonate or phosphonate prodrug can havethe structure A³ as described herein.

Stereoisomers

The compounds of the invention may have chiral centers, e.g., chiralcarbon or phosphorus atoms. The compounds of the invention thus includeracemic mixtures of all stereoisomers, including enantiomers,diastereomers, and atropisomers. In addition, the compounds of theinvention include enriched or resolved optical isomers at any or allasymmetric, chiral atoms. In other words, the chiral centers apparentfrom the depictions are provided as the chiral isomers or racemicmixtures. Both racemic and diastereomeric mixtures, as well as theindividual optical isomers isolated or synthesized, substantially freeof their enantiomeric or diastereomeric partners, are all within thescope of the invention. The racemic mixtures are separated into theirindividual, substantially optically pure isomers through well-knowntechniques such as, for example, the separation of diastereomeric saltsformed with optically active adjuncts, e.g., acids or bases followed byconversion back to the optically active substances. In most instances,the desired optical isomer is synthesized by means of stereospecificreactions, beginning with the appropriate stereoisomer of the desiredstarting material.

The compounds of the invention can also exist as tautomeric isomers incertain cases. Although only one delocalized resonance structure may bedepicted, all such forms are contemplated within the scope of theinvention. For example, ene-amine tautomers can exist for purine,pyrimidine, imidazole, guanidine, amidine, and tetrazole systems and alltheir possible tautomeric forms are within the scope of the invention.

Salts and Hydrates

Examples of physiologically acceptable salts of the compounds of theinvention include salts derived from an appropriate base, such as analkali metal (for example, sodium), an alkaline earth (for example,magnesium), ammonium and NX₄ ⁺ (wherein X is C₁-C₄ alkyl).Physiologically acceptable salts of a hydrogen atom or an amino groupinclude salts of organic carboxylic acids such as acetic, benzoic,lactic, fumaric, tartaric, maleic, malonic, malic, isethionic,lactobionic and succinic acids; organic sulfonic acids, such asmethanesulfonic, ethanesulfonic, benzenesulfonic and p-toluenesulfonicacids; and inorganic acids, such as hydrochloric, sulfuric, phosphoricand sulfamic acids. Physiologically acceptable salts of a compound of anhydroxy group include the anion of said compound in combination with asuitable cation such as Na⁺ and NX₄ ⁺ (wherein X is independentlyselected from H or a C₁-C₄ alkyl group).

For therapeutic use, salts of active ingredients of the compounds of theinvention will typically be physiologically acceptable, i.e. they willbe salts derived from a physiologically acceptable acid or base.However, salts of acids or bases which are not physiologicallyacceptable may also find use, for example, in the preparation orpurification of a physiologically acceptable compound. All salts,whether or not derived form a physiologically acceptable acid or base,are within the scope of the present invention.

Metal salts typically are prepared by reacting the metal hydroxide witha compound of this invention. Examples of metal salts which are preparedin this way are salts containing Li⁺, Na⁺, and K⁺. A less soluble metalsalt can be precipitated from the solution of a more soluble salt byaddition of the suitable metal compound.

In addition, salts may be formed from acid addition of certain organicand inorganic acids, e.g., HCl, HBr, H₂SO₄, H₃PO₄ or organic sulfonicacids, to basic centers, typically amines, or to acidic groups. Finally,it is to be understood that the compositions herein comprise compoundsof the invention in their un-ionized, as well as zwitterionic form, andcombinations with stoichiometric amounts of water as in hydrates.

Also included within the scope of this invention are the salts of theparental compounds with one or more amino acids. Any of the natural orunnatural amino acids are suitable, especially the naturally-occurringamino acids found as protein components, although the amino acidtypically is one bearing a side chain with a basic or acidic group,e.g., lysine, arginine or glutamic acid, or a neutral group such asglycine, serine, threonine, alanine, isoleucine, or leucine.

Methods of Inhibition of HCV

Another aspect of the invention relates to methods of inhibiting theactivity of HCV comprising the step of treating a sample suspected ofcontaining HCV with a compound or composition of the invention.

Compounds of the invention may act as inhibitors of HCV, asintermediates for such inhibitors or have other utilities as describedbelow. The inhibitors will generally bind to locations on the surface orin a cavity of the liver. Compounds binding in the liver may bind withvarying degrees of reversibility. Those compounds binding substantiallyirreversibly are ideal candidates for use in this method of theinvention. Once labeled, the substantially irreversibly bindingcompounds are useful as probes for the detection of HCV. Accordingly,the invention relates to methods of detecting NS3 in a sample suspectedof containing HCV comprising the steps of: treating a sample suspectedof containing HCV with a composition comprising a compound of theinvention bound to a label; and observing the effect of the sample onthe activity of the label. Suitable labels are well known in thediagnostics field and include stable free radicals, fluorophores,radioisotopes, enzymes, chemiluminescent groups and chromogens. Thecompounds herein are labeled in conventional fashion using functionalgroups such as hydroxyl or amino. In one embodiment the inventionprovides a compound of formula (I) that comprises or that is bound orlinked to one or more detectable labels. Within the context of theinvention samples suspected of containing HCV include natural orman-made materials such as living organisms; tissue or cell cultures;biological samples such as biological material samples (blood, serum,urine, cerebrospinal fluid, tears, sputum, saliva, tissue samples, andthe like); laboratory samples; food, water, or air samples; bioproductsamples such as extracts of cells, particularly recombinant cellssynthesizing a desired glycoprotein; and the like. Typically the samplewill be suspected of containing HCV. Samples can be contained in anymedium including water and organic solvent/water mixtures. Samplesinclude living organisms such as humans, and man made materials such ascell cultures.

The treating step of the invention comprises adding the compound of theinvention to the sample or it comprises adding a precursor of thecomposition to the sample. The addition step comprises any method ofadministration as described above.

If desired, the activity of HCV after application of the compound can beobserved by any method including direct and indirect methods ofdetecting HCV activity. Quantitative, qualitative, and semiquantitativemethods of determining HCV activity are all contemplated. Typically oneof the screening methods described above are applied, however, any othermethod such as observation of the physiological properties of a livingorganism are also applicable.

Many organisms contain HCV. The compounds of this invention are usefulin the treatment or prophylaxis of conditions associated with HCVactivation in animals or in man.

However, in screening compounds capable of inhibiting HCV it should bekept in mind that the results of enzyme assays may not always correlatewith cell culture assays. Thus, a cell based assay should typically bethe primary screening tool.

Screens for HCV Inhibitors

Compounds of the invention are screened for inhibitory activity againstHCV by any of the conventional techniques for evaluating enzymeactivity. Within the context of the invention, typically compounds arefirst screened for inhibition of HCV in vitro and compounds showinginhibitory activity are then screened for activity in vivo. Compoundshaving in vitro Ki (inhibitory constants) of less then about 5×10⁻⁶ M,typically less than about 1×10⁻⁷ M and preferably less than about 5×10⁻⁸M are preferred for in vivo use.

Useful in vitro screens have been described in detail.

Pharmaceutical Formulations

The compounds of this invention are formulated with conventionalcarriers and excipients, which will be selected in accord with ordinarypractice. Tablets will contain excipients, glidants, fillers, bindersand the like. Aqueous formulations are prepared in sterile form, andwhen intended for delivery by other than oral administration generallywill be isotonic. All formulations will optionally contain excipientssuch as those set forth in the Handbook of Pharmaceutical Excipients(1986). Excipients include ascorbic acid and other antioxidants,chelating agents such as EDTA, carbohydrates such as dextrin,hydroxyalkylcellulose, hydroxyalkylmethylcellulose, stearic acid and thelike. The pH of the formulations ranges from about 3 to about 11, but isordinarily about 7 to 10.

While it is possible for the active ingredients to be administered aloneit may be preferable to present them as pharmaceutical formulations. Theformulations, both for veterinary and for human use, of the inventioncomprise at least one active ingredient, as above defined, together withone or more acceptable carriers therefor and optionally othertherapeutic ingredients. The carrier(s) must be “acceptable” in thesense of being compatible with the other ingredients of the formulationand physiologically innocuous to the recipient thereof.

The formulations include those suitable for the foregoing administrationroutes. The formulations may conveniently be presented in unit dosageform and may be prepared by any of the methods well known in the art ofpharmacy. Techniques and formulations generally are found in Remington'sPharmaceutical Sciences (Mack Publishing Co., Easton, Pa.). Such methodsinclude the step of bringing into association the active ingredient withthe carrier which constitutes one or more accessory ingredients. Ingeneral the formulations are prepared by uniformly and intimatelybringing into association the active ingredient with liquid carriers orfinely divided solid carriers or both, and then, if necessary, shapingthe product.

Formulations of the present invention suitable for oral administrationmay be presented as discrete units such as capsules, cachets or tabletseach containing a predetermined amount of the active ingredient; as apowder or granules; as a solution or a suspension in an aqueous ornon-aqueous liquid; or as an oil-in-water liquid emulsion or awater-in-oil liquid emulsion. The active ingredient may also beadministered as a bolus, electuary or paste.

A tablet is made by compression or molding, optionally with one or moreaccessory ingredients. Compressed tablets may be prepared by compressingin a suitable machine the active ingredient in a free-flowing form suchas a powder or granules, optionally mixed with a binder, lubricant,inert diluent, preservative, surface active or dispersing agent. Moldedtablets may be made by molding in a suitable machine a mixture of thepowdered active ingredient moistened with an inert liquid diluent. Thetablets may optionally be coated or scored and optionally are formulatedso as to provide slow or controlled release of the active ingredienttherefrom.

For administration to the eye or other external tissues e.g., mouth andskin, the formulations are preferably applied as a topical ointment orcream containing the active ingredient(s) in an amount of, for example,0.075 to 20% w/w (including active ingredient(s) in a range between 0.1%and 20% in increments of 0.1% w/w such as 0.6% w/w, 0.7% w/w, etc.),preferably 0.2 to 15% w/w and most preferably 0.5 to 10% w/w. Whenformulated in an ointment, the active ingredients may be employed witheither a paraffinic or a water-miscible ointment base. Alternatively,the active ingredients may be formulated in a cream with an oil-in-watercream base.

If desired, the aqueous phase of the cream base may include, forexample, at least 30% w/w of a polyhydric alcohol, i.e. an alcoholhaving two or more hydroxyl groups such as propylene glycol, butane1,3-diol, mannitol, sorbitol, glycerol and polyethylene glycol(including PEG 400) and mixtures thereof. The topical formulations maydesirably include a compound which enhances absorption or penetration ofthe active ingredient through the skin or other affected areas. Examplesof such dermal penetration enhancers include dimethyl sulphoxide andrelated analogs.

The oily phase of the emulsions of this invention may be constitutedfrom known ingredients in a known manner. While the phase may comprisemerely an emulsifier (otherwise known as an emulgent), it desirablycomprises a mixture of at least one emulsifier with a fat or an oil orwith both a fat and an oil. Preferably, a hydrophilic emulsifier isincluded together with a lipophilic emulsifier which acts as astabilizer. It is also preferred to include both an oil and a fat.Together, the emulsifier(s) with or without stabilizer(s) make up theso-called emulsifying wax, and the wax together with the oil and fatmake up the so-called emulsifying ointment base which forms the oilydispersed phase of the cream formulations.

Emulgents and emulsion stabilizers suitable for use in the formulationof the invention include Tween® 60, Span® 80, cetostearyl alcohol,benzyl alcohol, myristyl alcohol, glyceryl mono-stearate and sodiumlauryl sulfate.

The choice of suitable oils or fats for the formulation is based onachieving the desired cosmetic properties. The cream should preferablybe a non-greasy, non-staining and washable product with suitableconsistency to avoid leakage from tubes or other containers. Straight orbranched chain, mono- or dibasic alkyl esters such as di-isoadipate,isocetyl stearate, propylene glycol diester of coconut fatty acids,isopropyl myristate, decyl oleate, isopropyl palmitate, butyl stearate,2-ethylhexyl palmitate or a blend of branched chain esters known asCrodamol CAP may be used, the last three being preferred esters. Thesemay be used alone or in combination depending on the propertiesrequired. Alternatively, high melting point lipids such as white softparaffin and/or liquid paraffin or other mineral oils are used.

Pharmaceutical formulations according to the present invention compriseone or more compounds of the invention together with one or morepharmaceutically acceptable carriers or excipients and optionally othertherapeutic agents. Pharmaceutical formulations containing the activeingredient may be in any form suitable for the intended method ofadministration. When used for oral use for example, tablets, troches,lozenges, aqueous or oil suspensions, dispersible powders or granules,emulsions, hard or soft capsules, syrups or elixirs may be prepared.Compositions intended for oral use may be prepared according to anymethod known to the art for the manufacture of pharmaceuticalcompositions and such compositions may contain one or more agentsincluding sweetening agents, flavoring agents, coloring agents andpreserving agents, in order to provide a palatable preparation. Tabletscontaining the active ingredient in admixture with non-toxicpharmaceutically acceptable excipients which are suitable formanufacture of tablets are acceptable. These excipients may be, forexample, inert diluents, such as calcium or sodium carbonate, lactose,lactose monohydrate, croscarmellose sodium, povidone, calcium or sodiumphosphate; granulating and disintegrating agents, such as maize starch,or alginic acid; binding agents, such as cellulose, microcrystallinecellulose, starch, gelatin or acacia; and lubricating agents, such asmagnesium stearate, stearic acid or talc. Tablets may be uncoated or maybe coated by known techniques including microencapsulation to delaydisintegration and adsorption in the gastrointestinal tract and therebyprovide a sustained action over a longer period. For example, a timedelay material such as glyceryl monostearate or glyceryl distearatealone or with a wax may be employed.

Formulations for oral use may be also presented as hard gelatin capsuleswhere the active ingredient is mixed with an inert solid diluent, forexample calcium phosphate or kaolin, or as soft gelatin capsules whereinthe active ingredient is mixed with water or an oil medium, such aspeanut oil, liquid paraffin or olive oil.

Aqueous suspensions of the invention contain the active materials inadmixture with excipients suitable for the manufacture of aqueoussuspensions. Such excipients include a suspending agent, such as sodiumcarboxymethylcellulose, methylcellulose, hydroxypropyl methylcelluose,sodium alginate, polyvinylpyrrolidone, gum tragacanth and gum acacia,and dispersing or wetting agents such as a naturally occurringphosphatide (e.g., lecithin), a condensation product of an alkyleneoxide with a fatty acid (e.g., polyoxyethylene stearate), a condensationproduct of ethylene oxide with a long chain aliphatic alcohol (e.g.,heptadecaethyleneoxycetanol), a condensation product of ethylene oxidewith a partial ester derived from a fatty acid and a hexitol anhydride(e.g., polyoxyethylene sorbitan monooleate). The aqueous suspension mayalso contain one or more preservatives such as ethyl or n-propylp-hydroxy-benzoate, one or more coloring agents, one or more flavoringagents and one or more sweetening agents, such as sucrose or saccharin.

Oil suspensions may be formulated by suspending the active ingredient ina vegetable oil, such as arachis oil, olive oil, sesame oil or coconutoil, or in a mineral oil such as liquid paraffin. The oral suspensionsmay contain a thickening agent, such as beeswax, hard paraffin or cetylalcohol. Sweetening agents, such as those set forth above, and flavoringagents may be added to provide a palatable oral preparation. Thesecompositions may be preserved by the addition of an antioxidant such asascorbic acid.

Dispersible powders and granules of the invention suitable forpreparation of an aqueous suspension by the addition of water providethe active ingredient in admixture with a dispersing or wetting agent, asuspending agent, and one or more preservatives. Suitable dispersing orwetting agents and suspending agents are exemplified by those disclosedabove. Additional excipients, for example sweetening, flavoring andcoloring agents, may also be present.

The pharmaceutical compositions of the invention may also be in the formof oil-in-water emulsions. The oily phase may be a vegetable oil, suchas olive oil or arachis oil, a mineral oil, such as liquid paraffin, ora mixture of these. Suitable emulsifying agents includenaturally-occurring gums, such as gum acacia and gum tragacanth,naturally occurring phosphatides, such as soybean lecithin, esters orpartial esters derived from fatty acids and hexitol anhydrides, such assorbitan monooleate, and condensation products of these partial esterswith ethylene oxide, such as polyoxyethylene sorbitan monooleate. Theemulsion may also contain sweetening and flavoring agents. Syrups andelixirs may be formulated with sweetening agents, such as glycerol,sorbitol or sucrose. Such formulations may also contain a demulcent, apreservative, a flavoring or a coloring agent.

The pharmaceutical compositions of the invention may be in the form of asterile injectable preparation, such as a sterile injectable aqueous oroleaginous suspension. This suspension may be formulated according tothe known art using those suitable dispersing or wetting agents andsuspending agents which have been mentioned above. The sterileinjectable preparation may also be a sterile injectable solution orsuspension in a non-toxic parenterally acceptable diluent or solvent,such as a solution in 1,3-butane-diol or prepared as a lyophilizedpowder. Among the acceptable vehicles and solvents that may be employedare water, Ringer's solution and isotonic sodium chloride solution. Inaddition, sterile fixed oils may conventionally be employed as a solventor suspending medium. For this purpose any bland fixed oil may beemployed including synthetic mono- or diglycerides. In addition, fattyacids such as oleic acid may likewise be used in the preparation ofinjectables.

The amount of active ingredient that may be combined with the carriermaterial to produce a single dosage form will vary depending upon thehost treated and the particular mode of administration. For example, atime-release formulation intended for oral administration to humans maycontain approximately 1 to 1000 mg of active material compounded with anappropriate and convenient amount of carrier material which may varyfrom about 5 to about 95% of the total compositions (weight:weight). Thepharmaceutical composition can be prepared to provide easily measurableamounts for administration. For example, an aqueous solution intendedfor intravenous infusion may contain from about 3 to 500 μg of theactive ingredient per milliliter of solution in order that infusion of asuitable volume at a rate of about 30 mL/hr can occur.

Formulations suitable for administration to the eye include eye dropswherein the active ingredient is dissolved or suspended in a suitablecarrier, especially an aqueous solvent for the active ingredient. Theactive ingredient is preferably present in such formulations in aconcentration of 0.5 to 20%, advantageously 0.5 to 10% particularlyabout 1.5% w/w.

Formulations suitable for topical administration in the mouth includelozenges comprising the active ingredient in a flavored basis, usuallysucrose and acacia or tragacanth; pastilles comprising the activeingredient in an inert basis such as gelatin and glycerin, or sucroseand acacia; and mouthwashes comprising the active ingredient in asuitable liquid carrier.

Formulations for rectal administration may be presented as a suppositorywith a suitable base comprising for example cocoa butter or asalicylate.

Formulations suitable for intrapulmonary or nasal administration have aparticle size for example in the range of 0.1 to 500 microns (includingparticle sizes in a range between 0.1 and 500 microns in incrementsmicrons such as 0.5, 1, 30 microns, 35 microns, etc.), which isadministered by rapid inhalation through the nasal passage or byinhalation through the mouth so as to reach the alveolar sacs. Suitableformulations include aqueous or oily solutions of the active ingredient.Formulations suitable for aerosol or dry powder administration may beprepared according to conventional methods and may be delivered withother therapeutic agents such as compounds heretofore used in thetreatment or prophylaxis of conditions associated with HCV activity.

Formulations suitable for vaginal administration may be presented aspessaries, tampons, creams, gels, pastes, foams or spray formulationscontaining in addition to the active ingredient such carriers as areknown in the art to be appropriate.

Formulations suitable for parenteral administration include aqueous andnon-aqueous sterile injection solutions which may contain anti-oxidants,buffers, bacteriostats and solutes which render the formulation isotonicwith the blood of the intended recipient; and aqueous and non-aqueoussterile suspensions which may include suspending agents and thickeningagents.

The formulations are presented in unit-dose or multi-dose containers,for example sealed ampoules and vials, and may be stored in afreeze-dried (lyophilized) condition requiring only the addition of thesterile liquid carrier, for example water for injection, immediatelyprior to use. Extemporaneous injection solutions and suspensions areprepared from sterile powders, granules and tablets of the kindpreviously described. Preferred unit dosage formulations are thosecontaining a daily dose or unit daily sub-dose, as herein above recited,or an appropriate fraction thereof, of the active ingredient.

It should be understood that in addition to the ingredients particularlymentioned above the formulations of this invention may include otheragents conventional in the art having regard to the type of formulationin question, for example those suitable for oral administration mayinclude flavoring agents.

The invention further provides veterinary compositions comprising atleast one active ingredient as above defined together with a veterinarycarrier therefor.

Veterinary carriers are materials useful for the purpose ofadministering the composition and may be solid, liquid or gaseousmaterials which are otherwise inert or acceptable in the veterinary artand are compatible with the active ingredient. These veterinarycompositions may be administered orally, parenterally or by any otherdesired route.

Compounds of the invention can also be formulated to provide controlledrelease of the active ingredient to allow less frequent dosing or toimprove the pharmacokinetic or toxicity profile of the activeingredient. Accordingly, the invention also provided compositionscomprising one or more compounds of the invention formulated forsustained or controlled release.

Effective dose of active ingredient depends at least on the nature ofthe condition being treated, toxicity, whether the compound is beingused prophylactically (lower doses), the method of delivery, and thepharmaceutical formulation, and will be determined by the clinicianusing conventional dose escalation studies. It can be expected to befrom about 0.0001 to about 100 mg/kg body weight per day. Typically,from about 0.01 to about 10 mg/kg body weight per day. More typically,from about 0.01 to about 5 mg/kg body weight per day. More typically,from about 0.05 to about 0.5 mg/kg body weight per day. For example, thedaily candidate dose for an adult human of approximately 70 kg bodyweight will range from 1 mg to 1000 mg, preferably between 5 mg and 500mg, and may take the form of single or multiple doses.

Routes of Administration

One or more compounds of the invention (herein referred to as the activeingredients) are administered by any route appropriate to the conditionto be treated. Suitable routes include oral, rectal, nasal, topical(including buccal and sublingual), vaginal and parenteral (includingsubcutaneous, intramuscular, intravenous, intradermal, intrathecal andepidural), and the like. It will be appreciated that the preferred routemay vary with for example the condition of the recipient. An advantageof the compounds of this invention is that they are orally bioavailableand can be dosed orally.

Combination Therapy

Active ingredients of the invention can also be used in combination withother active ingredients. Such combinations are selected based on thecondition to be treated, cross-reactivities of ingredients andpharmaco-properties of the combination.

It is also possible to combine any compound of the invention with one ormore other active ingredients in a unitary dosage form for simultaneousor sequential administration to a patient. The combination therapy maybe administered as a simultaneous or sequential regimen. Whenadministered sequentially, the combination may be administered in two ormore administrations.

The combination therapy may provide “synergy” and “synergistic effect”,i.e. the effect achieved when the active ingredients used together isgreater than the sum of the effects that results from using thecompounds separately. A synergistic effect may be attained when theactive ingredients are: (1) co-formulated and administered or deliveredsimultaneously in a combined formulation; (2) delivered by alternationor in parallel as separate formulations; or (3) by some other regimen.When delivered in alternation therapy, a synergistic effect may beattained when the compounds are administered or delivered sequentially,e.g., in separate tablets, pills or capsules, or by different injectionsin separate syringes. In general, during alternation therapy, aneffective dosage of each active ingredient is administered sequentially,i.e. serially, whereas in combination therapy, effective dosages of twoor more active ingredients are administered together.

Metabolites of the Compounds of the Invention

Also falling within the scope of this invention are the in vivometabolic products of the compounds described herein. Such products mayresult for example from the oxidation, reduction, hydrolysis, amidation,esterification and the like of the administered compound, primarily dueto enzymatic processes. Accordingly, the invention includes compoundsproduced by a process comprising contacting a compound of this inventionwith a mammal for a period of time sufficient to yield a metabolicproduct thereof. Such products typically are identified by preparing aradiolabelled (e.g., C¹⁴ or H³) compound of the invention, administeringit parenterally in a detectable dose (e.g., greater than about 0.5mg/kg) to an animal such as rat, mouse, guinea pig, monkey, or to man,allowing sufficient time for metabolism to occur (typically about 30seconds to 30 hours) and isolating its conversion products from theurine, blood or other biological samples. These products are easilyisolated since they are labeled (others are isolated by the use ofantibodies capable of binding epitopes surviving in the metabolite). Themetabolite structures are determined in conventional fashion, e.g., byMS or NMR analysis. In general, analysis of metabolites is done in thesame way as conventional drug metabolism studies well-known to thoseskilled in the art. The conversion products, so long as they are nototherwise found in vivo, are useful in diagnostic assays for therapeuticdosing of the compounds of the invention even if they possess noHCV-inhibitory activity of their own.

Methods for determining stability of compounds in surrogategastrointestinal secretions are known. Compounds are defined herein asstable in the gastrointestinal tract where less than about 50 molepercent of the protected groups are deprotected in surrogate intestinalor gastric juice upon incubation for 1 hour at 37° C. Simply because thecompounds are stable to the gastrointestinal tract does not mean thatthey cannot be hydrolyzed in vivo. The phosphonate prodrugs of theinvention typically will be stable in the digestive system but aresubstantially hydrolyzed to the parental drug in the digestive lumen,liver or other metabolic organ, or within cells in general.

Exemplary Methods of Making the Compounds of the Invention.

The invention also relates to methods of making the compositions of theinvention. The compositions are prepared by any of the applicabletechniques of organic synthesis. Many such techniques are well known inthe art. However, many of the known techniques are elaborated inCompendium of Organic Synthetic Methods (John Wiley & Sons, New York),Vol. 1, Ian T. Harrison and Shuyen Harrison, 1971; Vol. 2, Ian T.Harrison and Shuyen Harrison, 1974; Vol. 3, Louis S. Hegedus and LeroyWade, 1977; Vol. 4, Leroy G. Wade, Jr., 1980; Vol. 5, Leroy G. Wade,Jr., 1984; and Vol. 6, Michael B. Smith; as well as March, J., AdvancedOrganic Chemistry, Third Edition, (John Wiley & Sons, New York, 1985),Comprehensive Organic Synthesis. Selectivity, Strategy & Efficiency inModern Organic Chemistry. In 9 Volumes, Barry M. Trost, Editor-in-Chief(Pergamon Press, New York, 1993 printing). Other methods suitable forpreparing compounds of the invention are described in InternationalPatent Application Publication Number WO 2006/020276.

A number of exemplary methods for the preparation of the compositions ofthe invention are provided below. These methods are intended toillustrate the nature of such preparations and are not intended to limitthe scope of applicable methods.

Generally, the reaction conditions such as temperature, reaction time,solvents, work-up procedures, and the like, will be those common in theart for the particular reaction to be performed. The cited referencematerial, together with material cited therein, contains detaileddescriptions of such conditions. Typically the temperatures will be−100° C. to 200° C., solvents will be aprotic or protic, and reactiontimes will be 10 seconds to 10 days. Work-up typically consists ofquenching any unreacted reagents followed by partition between awater/organic layer system (extraction) and separating the layercontaining the product.

Oxidation and reduction reactions are typically carried out attemperatures near room temperature (about 20° C.), although for metalhydride reductions frequently the temperature is reduced to 0° C. to−100° C., solvents are typically aprotic for reductions and may beeither protic or aprotic for oxidations. Reaction times are adjusted toachieve desired conversions.

Condensation reactions are typically carried out at temperatures nearroom temperature, although for non-equilibrating, kinetically controlledcondensations reduced temperatures (0° C. to −100° C.) are also common.Solvents can be either protic (common in equilibrating reactions) oraprotic (common in kinetically controlled reactions).

Standard synthetic techniques such as azeotropic removal of reactionby-products and use of anhydrous reaction conditions (e.g., inert gasenvironments) are common in the art and will be applied when applicable.

The terms “treated”, “treating”, “treatment”, and the like, when used inconnection with a chemical synthetic operation, mean contacting, mixing,reacting, allowing to react, bringing into contact, and other termscommon in the art for indicating that one or more chemical entities istreated in such a manner as to convert it to one or more other chemicalentities. This means that “treating compound one with compound two” issynonymous with “allowing compound one to react with compound two”,“contacting compound one with compound two”, “reacting compound one withcompound two”, and other expressions common in the art of organicsynthesis for reasonably indicating that compound one was “treated”,“reacted”, “allowed to react”, etc., with compound two. For example,treating indicates the reasonable and usual manner in which organicchemicals are allowed to react. Normal concentrations (0.01M to 10M,typically 0.1M to 1M), temperatures (−100° C. to 250° C., typically −78°C. to 150° C., more typically −78° C. to 100° C., still more typically0° C. to 100° C.), reaction vessels (typically glass, plastic, metal),solvents, pressures, atmospheres (typically air for oxygen and waterinsensitive reactions or nitrogen or argon for oxygen or watersensitive), etc., are intended unless otherwise indicated. The knowledgeof similar reactions known in the art of organic synthesis are used inselecting the conditions and apparatus for “treating” in a givenprocess. In particular, one of ordinary skill in the art of organicsynthesis selects conditions and apparatus reasonably expected tosuccessfully carry out the chemical reactions of the described processesbased on the knowledge in the art.

Modifications of each of the exemplary schemes and in the examples(hereafter “exemplary schemes”) leads to various analogs of the specificexemplary materials produce. The above-cited citations describingsuitable methods of organic synthesis are applicable to suchmodifications.

In each of the exemplary schemes it may be advantageous to separatereaction products from one another and/or from starting materials. Thedesired products of each step or series of steps is separated and/orpurified (hereinafter separated) to the desired degree of homogeneity bythe techniques common in the art. Typically such separations involvemultiphase extraction, crystallization from a solvent or solventmixture, distillation, sublimation, or chromatography. Chromatographycan involve any number of methods including, for example: reverse-phaseand normal phase; size exclusion; ion exchange; high, medium, and lowpressure liquid chromatography methods and apparatus; small scaleanalytical; simulated moving bed (SMB) and preparative thin or thicklayer chromatography, as well as techniques of small scale thin layerand flash chromatography.

Another class of separation methods involves treatment of a mixture witha reagent selected to bind to or render otherwise separable a desiredproduct, unreacted starting material, reaction by product, or the like.Such reagents include adsorbents or absorbents such as activated carbon,molecular sieves, ion exchange media, or the like. Alternatively, thereagents can be acids in the case of a basic material, bases in the caseof an acidic material, binding reagents such as antibodies, bindingproteins, selective chelators such as crown ethers, liquid/liquid ionextraction reagents (LIX), or the like.

Selection of appropriate methods of separation depends on the nature ofthe materials involved. For example, boiling point, and molecular weightin distillation and sublimation, presence or absence of polar functionalgroups in chromatography, stability of materials in acidic and basicmedia in multiphase extraction, and the like. One skilled in the artwill apply techniques most likely to achieve the desired separation.

A single stereoisomer, e.g., an enantiomer, substantially free of itsstereoisomer may be obtained by resolution of the racemic mixture usinga method such as formation of diastereomers using optically activeresolving agents (Stereochemistry of Carbon Compounds, (1962) by E. L.Eliel, McGraw Hill; Lochmuller, C. H., (1975) J. Chromatogr., 113:(3)283-302). Racemic mixtures of chiral compounds of the invention can beseparated and isolated by any suitable method, including: (1) formationof ionic, diastereomeric salts with chiral compounds and separation byfractional crystallization or other methods, (2) formation ofdiastereomeric compounds with chiral derivatizing reagents, separationof the diastereomers, and conversion to the pure stereoisomers, and (3)separation of the substantially pure or enriched stereoisomers directlyunder chiral conditions.

Under method (1), diastereomeric salts can be formed by reaction ofenantiomerically pure chiral bases such as brucine, quinine, ephedrine,strychnine, α-methyl-β-phenylethylamine (amphetamine), and the like withasymmetric compounds bearing acidic functionality, such as carboxylicacid and sulfonic acid. The diastereomeric salts may be induced toseparate by fractional crystallization or ionic chromatography. Forseparation of the optical isomers of amino compounds, addition of chiralcarboxylic or sulfonic acids, such as camphorsulfonic acid, tartaricacid, mandelic acid, or lactic acid can result in formation of thediastereomeric salts.

Alternatively, by method (2), the substrate to be resolved is reactedwith one enantiomer of a chiral compound to form a diastereomeric pair(Eliel, E. and Wilen, S. (1994) Stereochemistry of Organic Compounds,John Wiley & Sons, Inc., p. 322). Diastereomeric compounds can be formedby reacting asymmetric compounds with enantiomerically pure chiralderivatizing reagents, such as menthyl derivatives, followed byseparation of the diastereomers and hydrolysis to yield the free,enantiomerically enriched xanthene. A method of determining opticalpurity involves making chiral esters, such as a menthyl ester, e.g., (−)menthyl chloroformate in the presence of base, or Mosher ester,α-methoxy-α-(trifluoromethyl)phenyl acetate (Jacob III. (1982) J. Org.Chem. 47:4165), of the racemic mixture, and analyzing the NMR spectrumfor the presence of the two atropisomeric diastereomers. Stablediastereomers of atropisomeric compounds can be separated and isolatedby normal- and reverse-phase chromatography following methods forseparation of atropisomeric naphthyl-isoquinolines (Hoye, T., WO96/15111). By method (3), a racemic mixture of two enantiomers can beseparated by chromatography using a chiral stationary phase (ChiralLiquid Chromatography (1989) W. J. Lough, Ed. Chapman and Hall, NewYork; Okamoto, (1990) J. of Chromatogr. 513:375-378). Enriched orpurified enantiomers can be distinguished by methods used to distinguishother chiral molecules with asymmetric carbon atoms, such as opticalrotation and circular dichroism.

Specific Embodiments of the Invention

International Patent Application Publication Number WO 2006/020276relates to certain specific compounds. In one specific embodiment of theinvention, the compounds of the invention exclude the followingcompounds:

In another embodiment of the invention the compounds of the inventionexclude the following compound:

In another embodiment of the invention the compounds of the inventionexclude a compound of formula (X):

or a pharmaceutically acceptable salt, or prodrug thereof, wherein:

-   -   R_(p) is cyclopentyl or tert-butyl;    -   Z¹ is selected from the following structures:

-   -   R_(a) is methoxy;    -   R_(b) is H;    -   R_(c) is phenyl that is optionally substituted with one or more        F, Cl, Br, I, (C1-6)alkyl, or (C1-6)alkoxy; and    -   A³ has any of the values defined herein.

In another embodiment of the invention the compounds of the inventionexclude a compound of formula (X):

or a pharmaceutically acceptable salt, or prodrug thereof, wherein:

-   -   R_(p) is (C1-6)alkyl or (C3-6)cycloalkyl;    -   Z¹ is selected from the following structures:

-   -   R_(a) is methoxy;    -   R_(b) is H;    -   R_(c) is phenyl that is optionally substituted with one or more        F, Cl, Br, I, (C1-6)alkyl, or (C1-6)alkoxy; and        A³ has any of the values defined herein.

In another embodiment of the invention the compounds of the inventionexclude a compound of formula (X):

or a pharmaceutically acceptable salt, or prodrug thereof, wherein:

-   -   R_(p) is cyclopentyl or tert-butyl;

Z¹ is selected from the following structures:

-   -   R_(a) is (C1-6)alkoxy;    -   R_(b) is H;    -   R_(c) is phenyl that is optionally substituted with one or more        F, Cl, Br, I, (C1-6)alkyl, or (C1-6)alkoxy; and    -   A³ has any of the values defined herein.

In another embodiment of the invention the compounds of the inventionexclude a compound of formula (X):

or a pharmaceutically acceptable salt, or prodrug thereof, wherein:

-   -   R_(p) is (C1-6)alkyl or (C3-6)cycloalkyl;    -   Z¹ is selected from the following structures:

-   -   R_(a) is (C1-6)alkoxy;    -   R_(b) is H;

R_(c) is phenyl that is optionally substituted with one or more F, Cl,Br, I, (C1-6)alkyl, or (C1-6)alkoxy; and

-   -   A³ has any of the values defined herein.

In another embodiment of the invention the compounds of the inventionexclude a compound of formula (XI):

or a pharmaceutically acceptable salt, or prodrug thereof, wherein:

-   -   R_(p) is cyclopentyl or tert-butyl;

Z¹ is selected from the following structures:

-   -   R_(a) is methoxy;    -   R_(b) is H;    -   R_(c) is phenyl that is optionally substituted with one or more        F, Cl, Br, I, (C1-6)alkyl, or (C1-6)alkoxy; and    -   A³ has any of the values defined herein.

In another embodiment of the invention the compounds of the inventionexclude a compound of formula (XI):

or a pharmaceutically acceptable salt, or prodrug thereof, wherein:

R_(p) is (C1-6)alkyl or (C3-6)cycloalkyl;

-   -   Z¹ is selected from the following structures:

-   -   R_(a) is methoxy;    -   R_(b) is H;    -   R_(c) is phenyl that is optionally substituted with one or more        F, Cl, Br, I, (C1-6)alkyl, or (C1-6)alkoxy; and        A³ has any of the values defined herein.

In another embodiment of the invention the compounds of the inventionexclude a compound of formula (XI):

or a pharmaceutically acceptable salt, or prodrug thereof, wherein:

-   -   R_(p) is cyclopentyl or tert-butyl;    -   Z¹ is selected from the following structures:

-   -   R_(a) is (C1-6)alkoxy;    -   R_(b) is H;    -   R_(c) is phenyl that is optionally substituted with one or more        F, Cl, Br, I, (C1-6)alkyl, or (C1-6)alkoxy; and    -   A³ has any of the values defined herein.

In another embodiment of the invention the compounds of the inventionexclude a compound of formula (XI):

or a pharmaceutically acceptable salt, or prodrug thereof, wherein:

-   -   R_(p) is (C1-6)alkyl or (C3-6)cycloalkyl;    -   Z¹ is selected from the following structures:

-   -   R_(a) is (C1-6)alkoxy;    -   R_(b) is H;    -   R_(c) is phenyl that is optionally substituted with one or more        F, Cl, Br, I, (C1-6)alkyl, or (C1-6)alkoxy; and    -   A³ has any of the values defined herein.

In a specific embodiment of the invention Z¹ is selected from thefollowing structures:

In a specific embodiment of the invention Z¹ is:

In a specific embodiment of the invention Z¹ is selected from thefollowing structures:

In a specific embodiment the invention provides a compound of formula(II):

or a pharmaceutically acceptable salt, or prodrug thereof, wherein:wherein:

R_(f) is H, alkyl, alkenyl, alkynyl, aryl, heteroaryl, or cycloalkyl,which R_(f) is optionally substituted with one or more R_(g);

each R_(g) is independently halo, hydroxy, cyano, arylthio, cycloalkyl,aryl, heteroaryl, alkoxy, NR_(h)R_(i), —C(═O)NR_(h)R_(i), wherein eacharyl and heteroaryl is optionally substituted with one or more alkyl,halo, hydroxy, cyano, nitro, amino, alkoxy, haloalkyl, or haloalkoxy;and

each R_(h) and R_(i) is independently H, alkyl, or haloalkyl.

In a specific embodiment of the invention R_(f) is alkyl, alkenyl, oralkynyl, which R_(f) is substituted with aryl that is optionallysubstituted with one or more alkyl, halo, hydroxy, cyano, nitro, amino,alkoxy, alkoxycarbonyl, alkanoyloxy, haloalkyl, or haloalkoxy.

In a specific embodiment of the invention R_(f) is alkyl, which issubstituted with aryl that is optionally substituted with one or morealkyl, halo, hydroxy, cyano, nitro, amino, alkoxy, alkoxycarbonyl,alkanoyloxy, haloalkyl, or haloalkoxy.

In a specific embodiment of the invention R_(f) is (C1-6)alkylsubstituted with a phenyl ring that is optionally substituted with 1, 2,or 3 alkyl, halo, hydroxy, cyano, nitro, amino, alkoxy, alkoxycarbonyl,alkanoyloxy, haloalkyl, or haloalkoxy.

In a specific embodiment of the invention R_(f) is benzyl or phenethylthat is optionally substituted with 1, 2, or 3 alkyl, halo, hydroxy,cyano, nitro, amino, alkoxy, alkoxycarbonyl, alkanoyloxy, haloalkyl, orhaloalkoxy.

In a specific embodiment of the invention R_(f) is H, methyl, ethyl,propyl, butyl, cyclopropylmethyl, 3-butenyl, 2-methylpropyl, isopropyl,vinyl, cis-1-propenyl, trans-1-propenyl, cis-1-butenyl,2-methylpropenyl, 2-phenylvinyl, 2-phenylethynyl, 3-methyl-2-butenyl,2-hydroxyethyl, 2-hydroxy-2-methylpropyl, cyanomethyl, methoxymethyl,N-(2,2,2-trifluoroethyl)-2-aminoethyl, phenethyl, 2-chlorophenethyl,2-fluorophenethyl, 2-methylphenethyl, 2-chloro-6-fluorophenethyl,phenylthiomethyl, benzyl, 4-fluorobenzyl, 3-fluorobenzyl,2-fluorobenzyl, 4-cyanobenzyl, 3-cyanobenzyl, 2-cyanobenzyl,4-methoxybenzyl, 3-methoxybenzyl, 2-methoxybenzyl, 2-bromobenzyl,2-trifluoromethoxybenzyl, 2-isopropoxybenzyl, 2-methylbenzyl,3-methylbenzyl, 4-methylbenzyl, 2-ethylbenzyl, 4-trifluoromethylbenzyl,3-trifluoromethylbenzyl, 2-trifluoromethylbenzyl, 4-chlorobenzyl,3-chlorobenzyl, 2-chlorobenzyl, 2,6-difluorobenzyl,2-chloro-6-fluorobenzyl, 2,6-dichlorobenzyl, 2-methoxy-6-fluorobenzyl,2,6-dimethylbenzyl, 2,6-difluoro-3-chlorobenzyl,2,6-difluoro-4-chlorobenzyl, 2-chloro-3,6-difluorobenzyl,2,3,6-trifluorobenzyl, 3-chloro-2,4-difluorobenzyl,2-chloro-3,6-difluorobenzyl, 2,3-dichloro-6-fluorobenzyl, 2-nitrobenzyl,2-aminobenzyl, 2-thienylmethyl, 2-furylmethyl, 3-furylmethyl,5-trifluoromethylfur-2-ylmethyl 5-pyrazolylmethyl, 2-oxazolylmethyl,4-methylthiazol-2-ylmethyl, 3-pyridyl, 2-pyridylmethyl,3-hydroxy-2-pyridylmethyl, 6-chloro-2-pyridylmethyl, 2-pyrazinylmethyl,5-pyrimidinylmethyl, 2-pyrimidinylmethyl, 4-pyrimidinylmethyl, phenyl,2-thiazolyl, N,N-dimethylaminocarbonylmethyl,N-methylaminocarbonylmethyl, aminocarbonylmethyl, 1-propynyl, or2-methylthiazol-4-ylmethyl.

In a specific embodiment the invention provides a compound of formula(III):

or a pharmaceutically acceptable salt, or prodrug thereof, wherein:

R_(j) is 1-[N-(2,2,2-trifluoroethyl)imino]ethyl, a,a-difluorophenethyl,cyclopropylacetyl, butanoyl, 4,4,4-trifluorobutanoyl,3,3,3-trifluoropropylsulfonyl, 3,3-dimethylbutanoyl,cyclopentylamino-carbonyl, cyclopropylacetyl, 2-norbornanylacetyl,2-amino-3,3-dimethylbutanoyl, 4-methylphenyl, 4-trifluoromethylphenyl,3-trifluoromethylphenyl, 2-trifluoromethylphenyl, or4-tert-butylthiazol-2-yl.

In a specific embodiment of the invention for a compound of formula(III), Z is O; Y¹ is O; and Z^(2a) and Z^(2b) are each hydrogen.

In a specific embodiment of the invention Q¹ is vinyl.

In a specific embodiment the invention provides a compound of formula(IV):

or a pharmaceutically acceptable salt, or prodrug thereof.

In a specific embodiment the invention provides a compound of formula(V):

or a pharmaceutically acceptable salt, or prodrug thereof.

In a specific embodiment the invention provides a compound of formula(VI):

or a pharmaceutically acceptable salt, or prodrug thereof, wherein:

R_(f) is H, alkyl, alkenyl, alkynyl, aryl, heteroaryl, or cycloalkyl,which R_(f) is optionally substituted with one or more R_(g);

each R_(g) is independently halo, hydroxy, cyano, arylthio, cycloalkyl,aryl, heteroaryl, alkoxy, NR_(h)R_(i), —C(═O)NR_(h)R_(i), wherein eacharyl and heteroaryl is optionally substituted with one or more alkyl,halo, hydroxy, cyano, nitro, amino, alkoxy, haloalkyl, or haloalkoxy;and

each R_(h), and R_(i) is independently H, alkyl, or haloalkyl; and

R_(j) is 1-[N-(2,2,2-trifluoroethyl)imino]ethyl, a,a-difluorophenethyl,cyclopropylacetyl, butanoyl, 4,4,4-trifluorobutanoyl,3,3,3-trifluoropropylsulfonyl, 3,3-dimethylbutanoyl,cyclopentylaminocarbonyl, cyclopropylacetyl, 2-norbornanylacetyl,2-amino-3,3-dimethylbutanoyl, 4-methylphenyl, 4-trifluoromethylphenyl,3-trifluoromethylphenyl, 2-trifluoromethylphenyl, or4-tert-butylthiazol-2-yl.

In a specific embodiment the invention provides a compound of formula(VII):

or a pharmaceutically acceptable salt, or prodrug thereof, wherein:

R_(f) is H, alkyl, alkenyl, alkynyl, aryl, heteroaryl, or cycloalkyl,which R_(f) is optionally substituted with one or more R_(g);

each R_(g) is independently halo, hydroxy, cyano, arylthio, cycloalkyl,aryl, heteroaryl, alkoxy, NR_(h)R_(i), —C(═O)NR_(h)R_(i), wherein eacharyl and heteroaryl is optionally substituted with one or more alkyl,halo, hydroxy, cyano, nitro, amino, alkoxy, haloalkyl, or haloalkoxy;and

each R_(h) and R_(i) is independently H, alkyl, or haloalkyl; and

R_(j) is 1-[N-(2,2,2-trifluoroethyl)imino]ethyl, a,a-difluorophenethyl,cyclopropylacetyl, butanoyl, 4,4,4-trifluorobutanoyl,3,3,3-trifluoropropylsulfonyl, 3,3-dimethylbutanoyl,cyclopentylaminocarbonyl, cyclopropylacetyl, 2-norbornanylacetyl,2-amino-3,3-dimethylbutanoyl, 4-methylphenyl, 4-trifluoromethylphenyl,3-trifluoromethylphenyl, 2-trifluoromethylphenyl, or4-tert-butylthiazol-2-yl.

In a specific embodiment the invention provides a compound which is aprodrug or a pharmaceutically acceptable salt thereof.

In a specific embodiment the invention provides a prodrug of formula(VIII):

or a pharmaceutically acceptable salt thereof, wherein:

R_(k) is a prodrug moiety.

In a specific embodiment of the invention R_(k) is benzyloxymethyl,pivaloyloxymethylcarbonate, 2-methylpropyloxy-carbonyloxymethyl,4-hydroxy-2-butenyl, benzoyloxymethyl, ethoxycarbonyloxymethyl, or agroup of the following formula:

In a specific embodiment the invention provides a compound of formula I,II, III, or VIII wherein Q¹ and Z^(2a) taken together with the atoms towhich they are attached form a 12-18 membered heterocycle, whichheterocycle may optionally be substituted with one or more oxo (═O) orA³.

In a specific embodiment the invention provides a compound of formula(IX):

or a pharmaceutically acceptable salt, or prodrug thereof.

In a specific embodiment the invention provides a compound of formula(X):

or a pharmaceutically acceptable salt, or prodrug thereof.

In a specific embodiment the invention provides a compound of formula(XI):

or a pharmaceutically acceptable salt, or prodrug thereof.

In a specific embodiment the invention provides a compound of formula(XII):

or a pharmaceutically acceptable salt, or prodrug thereof.

In a specific embodiment the invention provides a compound of formula(XIII):

or a pharmaceutically acceptable salt, or prodrug thereof.

In a specific embodiment the invention provides a compound of formula(XIV):

or a pharmaceutically acceptable salt, or prodrug thereof.

In a specific embodiment the invention provides a compound of formula(XV):

or a pharmaceutically acceptable salt, or prodrug thereof.

In a specific embodiment the invention provides a compound of formula(XVI):

or a pharmaceutically acceptable salt, or prodrug thereof.

In a specific embodiment the invention provides a compound of formula(XVII):

or a pharmaceutically acceptable salt, or prodrug thereof.

In a specific embodiment the invention provides a compound of formula(XVIII):

or a pharmaceutically acceptable salt, or prodrug thereof.

In a specific embodiment the invention provides a compound of formula(XXIV):

or a pharmaceutically acceptable salt, or prodrug thereof.

In a specific embodiment the invention provides a compound of formula(XXV):

or a pharmaceutically acceptable salt, or prodrug thereof.

In a specific embodiment the invention provides a compound of formula(XXVI):

or a pharmaceutically acceptable salt, or prodrug thereof.

In a specific embodiment the invention provides a compound of formula I:

or a pharmaceutically acceptable salt, or prodrug thereof, wherein:

-   -   R¹ is independently selected from H, alkyl, alkenyl, alkynyl,        aryl, cycloalkyl, heterocycle, halogen, haloalkyl,        alkylsulfonamido, arylsulfonamido, —C(O)NHS(O)₂—, or —S(O)₂—,        optionally substituted with one or more A³;    -   R² is selected from,    -   d) —C(Y¹)(A³),    -   e) (C2-10)alkyl, (C3-7)cycloalkyl or        (C1-4)alkyl-(C3-7)cycloalkyl, where said cycloalkyl and        alkyl-cycloalkyl may be optionally mono-, di- or tri-substituted        with (C1-3)alkyl, or        -   where said alkyl, cycloalkyl and alkyl-cycloalkyl may            optionally be mono- or di-substituted with substituents            selected from hydroxy and O—(C1-4)alkyl, or        -   where each of said alkyl-groups may optionally be mono-, di-            or tri-substituted with halogen, or        -   where each of said cycloalkyl groups being 5-, 6- or            7-membered, one or two —CH₂-groups not being directly linked            to each other may be optionally substituted replaced by —O—            such that the O-atom is linked to the N atom to which R² is            attached via at least two C-atoms,    -   f) phenyl, (C1-3)alkyl-phenyl, heteroaryl or        (C1-3)alkyl-heteroaryl, wherein the heteroaryl-groups are 5- or        6-membered having from 1 to 3 heteroatoms selected from N, O and        S, wherein said phenyl and heteroaryl groups may optionally be        mono-, di- or trisubstituted with substituents selected from        halogen, —OH, (C1-4)alkyl, O—(C1-4)alkyl, S—(C1-4)alkyl, —NH₂,        —CF₃, —NH((C1-4)alkyl) and —N((C1-4)alkyl)₂, —CONH₂ and        —CONH—(C1-4)alkyl; and wherein said (C1-3)alkyl may optionally        be substituted with one or more halogen; or    -   d) —S(O)₂(A³);    -   R₃ is H or (C1-6)alkyl;

Y¹ is independently O, S, N(A³), N(O)(A³), N(OA³), N(O)(OA³) orN(N(A³)(A³));

-   -   Z is O, S, or NR³;    -   Z¹ is selected from the following structures:

-   -   R_(a) is H or (C1-6)alkoxy;    -   R_(b) is H, F, Cl, Br, I, or (C1-6)alkyl;    -   R_(c) is H, cyano, F, Cl, Br, I, —C(═O)NR_(d)R_(e), or phenyl        that is optionally substituted with one or more F, Cl, Br, I,        (C1-6)alkyl, or (C1-6)alkoxy;    -   R_(d) and R_(e) are each independently H or (C1-6)alkyl;    -   each L is independently CH or N;    -   Z^(2a) is H, (C1-10)alkyl, (C2-10)alkenyl, (C2-10)alkynyl,        wherein any carbon atom may be replaced with a heteroatom        selected from O, S or N, or Z^(2a) optionally forms a carbocycle        or heterocycle with one or more R¹, R², Q¹, or A³;    -   Z^(2b) is H, (C1-6)alkyl, (C2-8)alkenyl, (C2-8)alkynyl;    -   Q¹ is (C1-8)alkyl, (C2-8)alkenyl, or (C2-8)alkynyl; or Q¹ and        Z^(2a) taken together with the atoms to which they are attached        form a carbocycle or heterocycle, which carbocycle or        heterocycle may optionally be substituted with one or more oxo        (═O) or A³;    -   A³ is independently selected from PRT, H, —OH, —C(O)OH, cyano,        alkyl, alkenyl, alkynyl, amino, amido, imido, imino, halogen,        CF₃, CH₂CF₃, cycloalkyl, nitro, aryl, aralkyl, alkoxy, aryloxy,        heterocycle, —C(A²)₃, —C(A²)₂-C(O)A², —C(O)A², —C(O)OA², —O(A²),        —N(A²)₂, —S(A²), —CH₂P(Y¹)(A²)(OA²), —CH₂P(Y¹)(A²)(N(A²)₂),        —CH₂P(Y¹)(OA²)(OA²), —OCH₂P(Y¹)(OA²)(OA²), —OCH₂P(Y¹)(A²)(OA²),        —OCH₂P(Y¹)(A²)(N(A²)₂), —C(O)OCH₂P(Y¹)(OA²)(OA²),        —C(O)OCH₂P(Y¹)(A²)(OA²), —C(O)OCH₂P(Y¹)(OA²)(N(A²)₂),        —CH₂P(Y¹)(OA²)(N(A²)₂), —OCH₂P(Y¹)(OA²)(N(A²)₂),        —C(O)OCH₂P(Y¹)(OA²)(N(A²)₂), —CH₂P(Y)(N(A²)₂)(N(A²)₂),        —C(O)OCH₂P(Y¹)(N(A²)₂)(N(A²)₂), —OCH₂P(Y¹)(N(A²)₂)(N(A²)₂),        —(CH₂)_(m)-heterocycle, —(CH₂)_(m)C(O)Oalkyl,        —O—(CH₂)_(m)—O—C(O)—Oalkyl, —O—(CH₂)_(r)—O—C(O)—(CH₂)_(m)-alkyl,        —(CH₂)_(m)O—C(O)—O-alkyl, —(CH₂)_(m)O—C(O)—O-cycloalkyl,        —N(H)C(Me)C(O)O-alkyl, or alkoxy arylsulfonamide,        -   wherein each A³ maybe optionally substituted with 1 to 4        -   R¹, —P(Y¹)(OA²)(OA²), —P(Y¹)(OA²)(N(A²)₂), —P(Y¹)(A²)(OA²),            —P(Y¹)(A²)(N(A²)₂), or P(Y¹)(N(A²)₂)(N(A²)₂), —C(═O)N(A²)₂),            halogen, alkyl, alkenyl, alkynyl, aryl, carbocycle,            heterocycle, aralkyl, aryl sulfonamide, aryl            alkylsulfonamide, aryloxy sulfonamide, aryloxy            alkylsulfonamide, aryloxy arylsulfonamide, alkyl            sulfonamide, alkyloxy sulfonamide, alkyloxy            alkylsulfonamide, arylthio, —(CH₂)_(m)heterocycle,            —(CH₂)_(m)—C(O)O-alkyl, —O(CH₂)_(m)OC(O)Oalkyl,            —O—(CH₂)_(m)—O—C(O)—(CH₂)_(m)-alkyl,            —(CH₂)_(m)—O—C(O)—O-alkyl, —(CH₂)_(m)—O—C(O)—O-cycloalkyl,            —N(H)C(CH₃)C(O)O-alkyl, or alkoxy arylsulfonamide,            optionally substituted with R¹;    -   Optionally each independent instance of A³ and Q¹ can be taken        together with one or more A³ or Q¹ groups to form a ring;    -   A² is independently selected from PRT, H, alkyl, alkenyl,        alkynyl, amino, amino acid, alkoxy, aryloxy, cyano, haloalkyl,        cycloalkyl, aryl, heteroaryl, alkylsulfonamide, or        arylsulfonamide, optionally substituted with A³; and    -   m is 0 to 6;        -   provided the compound is not a compound of any of the            following formulae:

SCHEMES AND EXAMPLES

General aspects of these exemplary methods are described below and inthe Examples. Each of the products of the following processes isoptionally separated, isolated, and/or purified prior to its use insubsequent processes.

A number of exemplary methods for the preparation of compounds of theinvention are provided herein, for example, in the Examples hereinbelow.These methods are intended to illustrate the nature of such preparationsare not intended to limit the scope of applicable methods. Certaincompounds of the invention can be used as intermediates for thepreparation of other compounds of the invention. For example, theinterconversion of various phosphonate compounds of the invention isillustrated below.

Preparation of Intermediates Preparation of Phosphonic AcidIntermediates 1. Synthesis and Resolution of Diethyl(1S,2R)-1-amino-2-ethenylcyclopropane-1-phosphonate dibenzoyl-L-tartaricAcid Salt

A solution of diethyl-(N-benzylideneaminomethyl)-phosphonate (50 g, 196mmol), trans-1,4-dibromo-2-butene (50 g, 235 mmol), andbenzyltriethylammonium chloride (4.5 g, 19.6 mmol) in dichloromethane(1.0 L) was stirred at rt using a mechanical stirrer when cesiumhydroxide monohydrate (82 g, 490 mmol) was added. The resulting mixturewas stirred for 18 hr after which another portion of cesium hydroxidemonohydrate (82 g, 490 mmol) was added. The resulting mixture wasstirred for 24 hr. The salts were then filtered off through a celite 521pad and the filtrate was allowed to stir with 1 N aq. HCl at rt for 3 h.The resulting mixture was filtered through another celite 521 pad andthe two phases of the filtrate were separated. The organic fraction wasextracted with 1 N aq. HCl (250 mL×1). The aqueous fractions were washedwith dichloromethane (250 mL×1) and the combined aq. fractions werestirred with ethyl acetate (500 mL) while 84 g (1 mol) of NaHCO₃ wasadded cautiously, followed by excess NaCl until saturated. After theresulting mixture was filtered through a celite 521 pad to remove excessNaCl and some black tar, the two layers were separated and the aqueousfraction was extracted further with ethyl acetate (250 mL×2). Theorganic extracts were washed with a saturated NaCl solution (250 mL×1),combined, dried (MgSO₄), and concentrated to obtain ˜16.5-17 g of thecrude amine.

The crude amine was partially purified by column chromatography using165-170 g of silica gel by eluting with ethyl acetate (100%, ˜500 mL),followed by 5% methanol in ethyl acetate (˜1200 mL). The productcontaining fractions were pooled and concentrated, which resulted11.5-12 g of partially purified amine.

To this amine was added a solution of 18.8-19.6 g (1 mole eq.) ofdibenzoyl-L-tartaric acid in 151.5-158 mL of acetonitrile (5 times theamount of the salt). The mixture was heated until it became a solutionand cooled slowly at rt to obtain solids. After overnight, the solidswere collected by filtration and washed with acetonitrile. The solidswere recrystallized from the same amount of acetonitrile again at rt toafford 11.5 g of optically pure salt.

¹H NMR (300 MHz, CD₃OD) d 8.14 (br, 2H), 8.11 (d, J=1.2 Hz, 2H), 7.64(tt, J=7.5 and 1.2 Hz, 2H), 7.51 (br t, J=7.5 Hz, 4H), 5.94 (s, 2H),5.82 (dt, J=17.1 and 9.9 Hz, 1H), 5.32 (dd, J=17.1 and 1.2 Hz, 1H), 5.13(dd, J=10.5 and 1.2 Hz, 1H), 4.11-4.26 (m, 4H), 2.11 (m, 1H), 1.33-1.47(m, 2H), 1.37 (dt, J=10.2 and 7.2 Hz, 6H); ³¹P NMR (121.4 MHz, CD₃OD) d22.55.

Analytical: The optical purity of the amine can be determined by ³¹P NMRof Mosher's amide in DMSO-d₆. The recrystallized material (25 mg) wasdissolved in a mixture of saturated aq. NaHCO₃ (5 mL) and saturated aq.NaCl (5 mL), and the free amine was extracted using dichloromethane (10mL×2). The extracts were washed once with a mixture of saturated aq.NaHCO₃ (5 mL) and saturated aq. NaCl (5 mL), dried (MgSO₄), andconcentrated.

To a solution of the residue and N,N-dimethylaminopyridine (˜3.5 mg) inpyridine (0.1 mL) was added(R)-(−)-a-methoxy-a-(trifluoromethyl)phenylacetyl chloride at rt. Afterstirring for 1.5 h, the pyridine was evaporated and the residue wasdissolved in 0.5 N HCl (10 mL) and ethyl acetate (10 mL). After theseparation of the two layers, the organic layer was washed with water(10 mL×1) and saturated aq. NaHCO₃ (10 mL×1), dried (MgSO₄), andconcentrated. In the ³¹P NMR of the residue in DMSO-d₆, the desiredamide appears at 23.00 ppm whereas the undesired amide comes at 22.79ppm.

2. Preparation of Phosphonic Acid Intermediates

Amine I (9.0 g, 41.1 mmol) was dissolved in 1,4-dioxane (100 mL). Asolution of Na₂CO₃ (13.1 g, 123.3 mmol) in H₂O (50 mL) was added to thereaction mixture and stirred for 5 minutes at rt. After benzylchloroformate (8.4 g, 49.3 mmol) was added, the reaction solution wasstirred at rt overnight. The organic phase was diluted with EtOAc andextracted with H₂O and brine. The organic phase was dried over MgSO₄.Concentration of the filtrate from vacuum filtration removal of theMgSO₄ yielded an oil from which II was isolated by column chromatography(SiO₂, 20% EtOAc in hexane) as a clear oil (11.6 g, 80%). ¹H NMR (300MHz, CDCl₃) d 7.33 (s, 5H), 6.05 (dt, J=9.9, 17.1 Hz, 1H), 5.65 (d,J=23.7 Hz, 1H), 5.³¹ (d, J=17.1 Hz, 1H), 5.06 (m, 3H), 4.06 (m, 4H),2.09 (m, 1H), 1.73 (m, 2H), 1.15 (dt, J=8.1, 26.4 Hz, 6H). ³¹P NMR(121.4 MHz, CDCl₃) d 23.7

Intermediate II (11.6 g, 32.9 mmol) and NaI (24.5 g, 164.3 mmol) weredissolved in pyridine (110 mL). The reaction solution was heated to 115°C. for 10 hours. After cooling back to rt, the reaction solution wasconcentrated to remove pyridine. H₂O (50 mL) was added to the crude. Theaqueous was washed by diethyl ether (2×100 mL). Then the aqueous phasewas adjusted to pH=2 by adding 1 M HCl_((aq.)). Product III (7.5 g, 23.0mmol) was isolated by extracting with dichloromethane and used for nextstep without further purification. ¹H NMR (300 MHz, CDCl₃) d 8.63 (br,1H), 7.33 (s, 5H), 5.95 (dt, J=9.9, 17.1 Hz, 1H), 5.65 (d, J=23.7 Hz,1H), 5.³¹ (d, J=17.1 Hz, 1H), 5.06 (m, 3H), 4.06 (m, 2H), 2.09 (m, 1H),1.73 (m, 2H), 1.23 (dt, J=8.1, 26.4 Hz, 3H)

³¹P NMR (121.4 MHz, CDCl₃) d 24.6. LC/MS=326 (M⁺+1), 348 (M⁺+Na)

3. Preparation of Phosphinic Acid Intermediates

A general scheme for the preparation of phosphinic acid is shown belowstarting from compound III (Scheme 1).

An alternative scheme (Scheme 2) for the preparation of phosphinic acidis shown below.

Phosphonic acid intermediate III (1.0 g, 3.1 mmol) was dissolved intoluene (6 mL). This solution was then added dropwise to (COCl)₂ (1.1mL, 12.4 mmol) and DMF (47 μL, 0.6 mmol) dissolved in 6 mL of toluene atrt. After 1 hour of stirring at rt, the reaction was concentrated andazeotroped three times with toluene to afford crude IV as an oil.

The resulting dark, viscous residue in THF (20 mL) was stirred at −78°C. as 1.0 M LiAlH(O-tBu)₃ (23.5 mL, 23.5 mmol) was added over 10minutes. The solution was warmed to r.t. over 30 minutes. The reactionmixture was cooled to 0° C. and quenched with ice cold 1 N HCl (200 mL).The product was extracted with ether (200 mL×2) and the organicfractions were washed with ice cold 1 N HCl (100 mL) and H₂O (100 mL).After the organic fraction was dried (MgSO₄) and concentrated, theresidue was purified by combi-flash column chromatography usinghexane/ethyl acetate as eluent to obtain IV (1.89 g, 78.3%). ¹H NMR (300MHz, CDCl₃): d 8.14 (bs, 1H), 7.35 (s, 5H), 6.22 (s, 1H), 5.89 (m, 2H),5.39 (bd, J=11.7 Hz, 1H), 5.30 (s, 2H), 5.21-5.104 (m, 3H), 4.13 (m,2H), 2.16 (m, 1H), 1.72-1.66 (m, 2H), 1.31 (m, 3H).

³¹P (121.4 MHz, CD₃OD):

32.311, 29.241

The resulting phosphinic acid is coupled with dipeptide intermediate asshown in Scheme 3.

4. Preparation of Dipeptide Intermediates A. Synthesis of PhenylQuinoline Dipeptide Intermediate

Step 1. Quinoline (7.6 g, 30.1 mmol), N-t-Boc-cis-4-hydroxy-L-prolinemethyl ester (8.9 g, 36.3 mmol) and triphenylphosphine (17.4 g, 66.3mmol) were dissolved in THF (250 mL). After cooling the reactionsolution to 0° C., DIAD (13.4 g, 66.3 mmol) was added over 15 minutes.The reaction was stirred at rt for 12 hours and was diluted with EtOAc(700 mL) and washed with NaHCO_(3 (aq.)), H₂O and brine. The organicphase was dried over MgSO₄. After concentration, a crystallization wasused to remove most of the triphenylphosphine oxide by using EtOAc (100mL) and hexane (50 mL) and desired product was isolated by columnchromatography (SiO₂, 70% EtOAc in hexane) as an oil (11.9 g, 85%). ¹HNMR (300 MHz, CDCl₃) d 8.03 (m, 2H), 7.50 (m, 5H), 7.18 (m, 1H), 6.97(m, 1H), 5.15 (m, 1H), 4.99 (m, 2H), 4.06 (s, 3H), 3.99 (m, 1H), 3.75(s, 3H), 2.79 (dd, J=8.7, 14.3 Hz, 1H), 2.45 (ddd, J=3.5, 10.7, 13.8 Hz,1H), 1.15 (s, 9H).

LC/MS=479 (M⁺+1), 501 (M⁺+Na)

Step 2. Product from the above reaction (9.6 g, 20.8 mmol) was dissolvedin dichloromethane (20 mL). 4.0 M HCl in 1,4-dioxane (50 mL) was addedto the reaction solution slowly and the reaction solution was allowed tostir at rt for 5 hours. After concentration under high vacuum for 30minutes, the crude was dissolved in DMF (70 mL). Acid (6.1 g, 25.0mmol), HATU (11.9 g, ³¹.2 mmol) and N-methylmorpholine (10.5 g, 104.0mmol) were added to the reaction solution. The reaction solution wasstirred at rt overnight and was diluted with EtOAc (500 mL) and washedwith NH₄Cl_((aq.)), NaHCO_(3(aq.)) and brine. The organic phase wasdried over MgSO₄. After concentration, the desired product (10.0 g, 80%)was isolated by column chromatography (SiO₂, 90% EtOAc in hexane) as asolid. ¹H NMR (300 MHz, CD₃OD) d 8.33 (d, J=9.6 Hz, 1H), 8.09 (m, 2H),7.74 (m, 3H), 7.65 (m 1H), 7.52 (m 1H), 7.24 (dd, J=2.1, 9.6 Hz, 1H),5.91 (m, 1H), 5.04 (m, 1H), 4.81 (d, J=9.0 Hz, 1H), 4.76 (d, J=9.0 Hz,1H), 4.46 (m, 1H), 4.23 (m, 1H), 4.06 (s, 3H), 3.99 (m, 1H), 3.75 (s,3H), 2.99 (dd, J=9.0, 14.7 Hz, 1H), 2.53 (ddd, J=3.3, 10.5, 13.8 Hz,1H), 1.42-1.78 (m, 8H), 1.05 (s, 9H). LC/MS=604 (M⁺+1), 626 (M⁺+Na).

Step 3. The methyl ester (9.2 g, 15.3 mmol) was dissolved in THF (30mL), MeOH (10 mL) and H₂O (10 mL). LiOH (1.8 g, 76.5 mmol) was added tothe reaction solution and the reaction solution was allowed to stir atrt for 7 hours. After EtOAc (150 mL) was added to dilute the reactionsolution, the aqueous phase was adjusted to pH=2 by adding 1 MHCl_((aq.)). Dipeptide acid VI (8.6 g, 95%) was isolated by extractingwith EtOAc (2×100 mL) and used for next step without furtherpurification. ¹H NMR (300 MHz, CD₃OD) d 8.38 (d, J=9.6 Hz, 1H), 8.11 (m,2H), 7.76 (m, 3H), 7.65 (m 1H), 7.55 (m 1H), 7.24 (dd, J=2.1, 9.6 Hz,1H), 5.89 (m, 1H), 5.04 (m, 1H), 4.81 (d, J=8.7 Hz, 1H), 4.76 (d, J=8.7Hz, 1H), 4.46 (m, 1H), 4.23 (m, 1H), 4.06 (s, 3H), 3.99 (m, 1H), 2.99(dd, J=9.0, 14.7 Hz, 1H), 2.53 (ddd, J=3.3, 10.5, 13.8 Hz, 1H),1.42-1.78 (m, 8H), 1.05 (s, 9H)

LC/MS=590 (M⁺+1), 612 (M⁺+Na).

B. Synthesis of1-(2-Cyclopentyloxycarbonylamino-3,3-dimethyl-butyryl)-4-[2-(2-isopropylamino-thiazol-4-yl)-7-methoxy-quinolin-4-yloxy]-pyrrolidine-2-carboxylicAcid

Step 1. To a solution of hydroxythiazole quinoline (20.0 g, 63.5 mmol)in THF (400 mL), was added cis-Boc-hydroxyproline methyl ester (18.7 g,76.2 mmol), and triphenylphosphine (36.6 g, 139.7 mmol). The solutionwas cooled to 0° C. and DIAD (27 mL, 139.7 mmol) was added slowly. Thesolution was allowed to warm to rt over a period of 1 h and stirredovernight. The solvent was removed under reduced pressure and the crudereaction mixture was dissolved in ethyl acetate and extracted with waterfollowed by brine. The organics were dried over MgSO₄, filtered and thesolvent was removed under reduced pressure. The crude material waseluted through a plug of silica using a quick gradient of (25%-100%)ethyl acetate/hexane to afford 32.5 g of desired product as a yellowsolid that has 10%-15% triphenylphosphineoxide contamination. ¹H NMR(300 MHz, CDCl₃): d 7.98, (d, J=9.2 Hz, 1H), 7.46 (m, 2H), 7.37 (d,J=2.4 Hz, 1H), 7.³¹ (s, 1H), 7.09 (d, J=9.1 Hz, 1H), 5.26 (m, 1H), 4.96(m, 1H), 4.62 (t, J=7.3 Hz, 1H), 5.57 (t, J=15 Hz, 1H), 3.97-3.84 (bs,5H), 3.76-3.66 (bs, 5H), 2.77 (m, 1H), 2.42 (m, 1H), 2.03 (s, 1H), 1.43(s, 9H), 1.33 (d, J=6.4 Hz, 6H). LC/MS: 543 (M⁺+1).

Step 2. To a solution of methyl ester (30.0 g, 55 mmol) in methylenechloride (150 mL) at 0° C., was added 4 N HCl in dioxane (150 mL). Thereaction was allowed to warm to rt over 1 hr. As the reaction proceeds,the product precipitates out of solution. The solids were filtered offand then washed repeatedly with diethyl ether to afford the HCl salt ofthe amine (20.67 g, 78%) as a crystalline yellow solid. ¹H NMR (300 MHz,CD₃OD): d 8.45 (d, J=9.2 Hz, 1H), 8.35 (s, 1H), 7.85 (s, 1H), 7.79 (s,1H), 7.45 (d, J=9.5 Hz, 1H), 6.02 (m, 1H), 4.22 (m, 1H), 4.07 (s, 3H),4.02 (d, J=3.9 Hz, 1H), 3.98 (s, 1H), 3.92 (s, 3H), 3.66 (s, 1H), 3.03(m, 1H), 2.82 (m, 1H), 1.36 (d, J=6.4 Hz, 6H), 1.33 (d, J=6.4 Hz, 6H).LC/MS: 443 (M⁺+1). To a solution of the HCl amine salt (20.96 g, 43.8mmol) in DMF (300 mL) at rt was added cyclopentylcarbamate-tert-leucinecarboxylic acid (13.0 g, 52.6 mmol), and HATU (25.0 g, 65.7 mmol). Thereaction was stirred for 10 min at rt and then Hunig's base (45 mL, 262mmol) was added over 5 min. The reaction was stirred at rt for 1 h,monitoring by LCMS. Solvent was removed under reduced pressure and theresidue was diluted with ethyl acetate. The reaction was extracted withsat. NaHCO₃, followed by water and brine. The organics were dried overMgSO₄, the solids were removed by filtration and then the solvent wasremoved under reduced pressure. The crude material was eluted through asilica plug to remove excess salts. The solvent was removed, and theproduct was recrystallized from ethyl acetate and hexane to afforddipeptide methyl ester (23.5 g, 81%) as a yellow crystalline solid. ¹HNMR (300 MHz, CDCl₃): d 7.98, (d, J=9.1 Hz, 1H), 7.67 (s, 1H), 7.51 (s,1H), 7.27 (s, 1H), 7.16 (d, J=7.3 Hz, 1H), 5.62 (m, 1H), 5.54 (m, 1H),5.27 (d, J=9.7 Hz, 1H), 4.81-4.71 (bs, 2H), 4.49 (d, J=12.5 Hz, 1H),4.28 (d, J=10 Hz, 1H), 4.14 (m, 1H), 4.04 (s, 3H), 3.78 (s, 3H), 3.60(m, 1H), 2.76 (m, 2H), 2.51 (m, 2H), 1.63-1.50 (m, 10H) 1.26 (d, J=6.4Hz, 6H), 1.07 (s, 9H). LC/MS: 668 (M⁺+1).

Step 3. To a solution of methyl ester (21.0 g, ³¹0.5 mmol) in THF (300mL) and methanol (15 mL) was added lithium hydroxide powder (4.5 g, 187mmol) in water (150 mL). The reaction was stirred at rt overnight. Theorganic solvents were removed under reduced pressure and adjusted to pH2-3 with 10% HCl in water. The solution was extracted with ethylacetate, (2×250 mL). The combined organics were dried over MgSO₄, whichwas removed by filtration, and the solvent was removed under reducedpressure to afford dipeptide carboxylic acid VII (19.3 g, 94%) as ayellow solid. ¹H NMR (300 MHz, CD₃OD): d 8.29 (d, J=9.5 Hz, 1H), 8.17(s, 1H), 7.72 (s, 2H), 7.33 (d, J=7.6 Hz, 1H), 5.77 (s, 1H), 4.80 (t,J=9.1 Hz, 1H), 4.77 (d, J=12 Hz, 1H), 4.44 (m, 1H), 4.19-4.04 (bs, 6H),2.96 (m, 1H), 2.50 (m, 1H), 1.62-1.50 (bs, 8H), 1.35 (d, J=6.7 Hz, 6H),1.05 (s, 9H). LC/MS: 655 (M⁺+1).

5. Preparation of Dipeptide Intermediates

Synthesis of dipeptide intermediates is shown in Scheme 4 and Scheme 5.

Amine (7.00 g, 28.55 mmol) and DABCO (5.13 g, 45.94 mmol) were dissolvedin toluene (30 mL). A toluene (11 mL) solution of brosylchloride (10.22g, 40.01 mmol) was added. The reaction mixture was stirred at rtovernight. The reaction was diluted with EtOAc (210 mL) and 0.5 N HCl(200 mL) was added. The two layers were separated and the aqueous layerwas extracted with EtOAc (2×200 mL). The combined organic layers werewashed with brine (200 mL), dried with Na₂SO₄, filtered, andconcentrated. The crude product was purified by combi-flash to give12.23 g of intermediate IX in 92% yield.

To a solution of X (12.8 g, 20.7 mmol) in CH₂Cl₂ (50 mL) was added 4 NHCl in 1,4-dioxane (50 mL, 200 mmol). The reaction mixture was stirredat r.t. for 2 h, concentrated, dried under vacuum for 20 minutes, andthen dissolved in CH₃CN (50 mL). Saturated NaHCO₃ in H₂O (50 mL) wasadded and stirred for 5 minutes. Freshly preparedcyclopentylchloroformate in THF (50 mL) was added. The reaction wascomplete within 1 h. The solvent was removed under reduced pressure andthe residue was diluted with EtOAc. The mixture was brought to pH=2 with1 N HCl and the two layers were separated. The organic layers werewashed with brine, dried with Na₂SO₄, filtered, and concentrated to givecrude product (3.18 g).

The crude ester (3.18 g, 5.07 mmol) was dissolved in THF (25 mL), H₂O(25 mL), and then MeOH (6 mL) and LiOH (660 mg, 25.4 mmol) was added.The reaction mixture was stirred at rt for 1 h and diluted with EtOAc.The reaction mixture was acidified to pH 2 with 1 N HCl and the twolayers were separated. The aqueous layer was extracted with EtOAc (2×).The combined organic layers were washed with brine, dried with Na₂SO₄,concentrated and dried under vacuum to give 3.09 g of acid XI.

The proline could be coupled to phosphinate to provide dipeptide asshown in Scheme 6.

6. Preparation of8-Chloro-2-(2-isopropylamino-thiazol-4-yl)-7-methoxy-quinolin-4-ol

The synthesis of 8-chloro quinoline is shown in scheme 7. The samesynthesis is used to prepare 8-bromo, fluoro and methyl analogs.

8-chloro-4-hydroxy-7-methoxyquinoline-2-carboxylic acid:

To a solution of methyl8-chloro-4-hydroxy-7-methoxyquinoline-2-carboxylate (36.5 g, 0.145 mol)in a mixture of 1:1 of MeOH: THF (160 mL total) was added a solution ofLiOH (30.5 g, 0.725 mol) in H₂O (80 mL). The mixture was stirred at roomtemperature for an hour when LCMS analysis showed complete conversion tothe carboxylic acid. The reaction was worked up by removal of thevolatiles and adjusting the pH of the solution to 6 using aqueous 6NHCl. The resulted gummy residue was filtered and dried on the lypholizerfor 2 days to provide 34.4 g (99.6%) of the product as a white solid. EIMS (m/z) 253.9 [M+H].

2-(2-diazo-1-oxo)-8-chloro-7-methoxyquinolin-4-yl isobutyl carbonate:

To a solution of 8-chloro-4-hydroxy-7-methoxyquinoline-2-carboxylic acid(10.2 g, 0.04 mol) in THF (400 mL) was added triethyl amine (12.3 mL,0.088 mol) and i-Butylchloroformate (11.6 mL, 0.088 mol) at 0° C. underan argon atmosphere. The mixture was stirred at 0° C. for 1 hour whenLCMS analysis demonstrated completion of the reaction to provide thedesired mixed anhydride. EI MS (m/z) 454.0 [M+H].

To the reaction mixture of the anhydride was added a 1M solution ofdiazomethane (121 mL, 0.121 mol) in diethyl ether via a plastic funnelat 0° C. This mixture was allowed to stir while warming up to roomtemperature for additional 2 hours. Analysis of the mixture by LCMSdemonstrated completion of the reaction. The septum was removed and thereaction was stirred for additional 20 minutes before removal of thesolvent. The residue resulted was dried further under high vacuum andcarried on to the next step. EI MS (m/z) 377.9 [M+H].

Preparation of Diazomethane from MNNG:

To a solution of 130 mL of 40% aqueous KOH and 130 mL of diethyl etheron ice was added a slurry of N-methyl-N′-nitro-N-nitrosoguanidine (18 g,0.121 mol) over 15 minutes. The mixture was stirred on ice foradditional 15 minutes when no further bubbling was observed. The organiclayer was decanted to another flask and stored over KOH pellets forsubsequent use.

8-chloro-2-(2-(isopropylamino)thiazol-4-yl)-7-methoxyquinolin-4-ol:

To a cooled solution of2-(2-diazo-1-oxo)-8-chloro-7-methoxyquinolin-4-yl isobutyl carbonate(15.2 g, 0.040 mol) at 0° C. in THF (268 mL) was added 48% HBr (23 mL,0.201 mol) slowly over 15 minutes. The solution was stirred at 0° C. foran additional 40 minutes when LCMS analysis demonstrated completereaction. The reaction was worked up by addition of aqueous 1N NaOH (180mL) at 0° C. to adjust the pH of the aqueous layer to 9. The layers wereseparated and the aqueous layer was washed with EtOAc (2×200 mL).Combined organic extracts were washed with brine and dried over MgSO₄.The solvent was removed in vacuo to provide 17.7 g of a yellow solid. EIMS (m/z) 4³¹.9 [M+H].

The solution of the bromoketone obtained from the previous reaction wassuspended in i-propanol (270 mL) and heated at 72° C. for 2 hours whenLCMS analysis of the reaction demonstrated complete conversion to thedesired product. The reaction was allowed to cool to room temperature toallow for the product to precipitate out of the solution. The reactionwas further cooled to 0° C. for 12 hours before filtration. The filtratewas washed with ether and dried on lypholizer to provide 8.03 g of thedesired product as an orange solid. ¹H NMR (500 MHz, CDCl₃): d 8.21 (d,J=9 Hz, 1H), 7.74 (s, 1H), 7.44 (d, J=10 Hz), 1H), 7.07 (s, 1H), 4.05(s, 3H), 3.92 (pentet, J=6 Hz, 1H), 1.25 (d, J=7 Hz, 6H). EI MS (m/z)350.0 [M+H].

Example 1 Preparation of Compound 1

Phosphonic acid intermediate(1-benzyloxycarbonylamino-2-vinyl-cyclopropyl)-phosphonic acid monoethylester III (415 mg, 1.28 mmol) was dissolved in toluene (8 mL). Thissolution was cooled to 0° C. and (COCl)₂ (222 μL, 2.56 mmol) was addedin a drop-wise fashion. DMF (44 μL, 0.56 mmol) was then added. Thereaction was run for 2 h at 0° C. and determined to be complete by ³¹PNMR. ³¹P NMR (121.4 MHz, CDCl₃): d 39.0, 38.5, 37.4, 36.5, 17.0, 16.2,16.0, 15.4.

The reaction was concentrated to orange-yellow oil and then placed underhigh vacuum for 1 h. The resulting residue was dissolved in THF (6.4 mL)and this solution was cooled to −78° C. A 1.4 M solution ofmethyllithium in diethyl ether (1.37 mL, 1.92 mmol) was added drop-wise.After 40 min, more methyllithium (456 μL, 0.64 mmol) was addeddrop-wise. After 10 min, the reaction was quenched at −78° C. by theaddition of sat. NH₄Cl_((aq.)). The organic phase was diluted with EtOAcand extracted with sat. NH₄Cl_((aq.)) and brine. The organic phase wasdried over MgSO₄. Concentration of the filtrate after removal of theMgSO₄ by vacuum filtration yielded an orange oil from which the productwas isolated by column chromatography (SiO₂, 100% EtOAc) as a clear oil(214 mg, 52% over 2 steps). ¹H NMR (300 MHz, CDCl₃): d 7.33 (s, 5H),6.09 (dt, J=9.9, 17.1 Hz, 1H), 5.65 (d, J=23.7 Hz, 1H), 5.³¹ (d, J=17.1Hz, 1H), 5.06 (m, 3H), 4.06 (m, 2H), 2.09 (m, 1H), 1.73 (m, 2H), 1.40(d, 3H), 1.13 (dt, J=8.1, 26.4 Hz, 3H). ³¹P NMR (121.4 MHz, CDCl₃): d53.7, 50.8. LC/MS=324 (M⁺+1), 346 (M⁺+Na)

A solution of(1-benzyloxycarbonylamino-2-vinyl-cyclopropyl)-methyl-phosphinic acidethyl ester (100 mg, 0.308 mmol) in CH₃CN (7.7 mL) was cooled to 0° C.and TMSI (220 μL, 1.54 mmol) was added in a drop-wise fashion. Thereaction was warmed to rt and stirred for an hour. The reaction was thencooled to 0° C. and additional TMSI (110 μL, 0.77 mmol) was added in adrop-wise fashion. The reaction was warmed to rt and stirred for 30 min.The reaction was cooled back to 0° C. and 2,6-lutidine (360 μL, 3.1mmol) was added in a drop-wise fashion. This was followed by theaddition of Et₃N (1 mL, 7.2 mmol) and MeOH (4 mL). The reaction was thenconcentrated in vacuo and the crude amino phosphinic acid was useddirectly in the next reaction.

A solution of dipeptide VII (81 mg, 0.123 mmol) in THF (2 mL) was cooledto −30° C. Et₃N (34 μL, 0.246 mmol) was added to this solution followedby ClCO₂Et (18 μL, 0.185 mmol). The reaction was stirred at atemperature between −20° C. and −30° C. for 30 min. Additional Et₃N (34μL, 0.246 mmol) and ClCO₂Et (18 μL, 0.185 mmol) were added. The reactionmixture was stirred for an additional 30 min at a temperature between−20° C. and −30° C. A solution of the crude amino phosphinic acid inCH₂Cl₂ (2 mL) was added in a drop-wise fashion at −30° C. and thereaction was warmed to rt and stirred for 2 hours. The reaction wasquenched by the addition of sat. NH₄Cl_((aq.)). The organic phase wasdiluted with EtOAc and washed sequentially with sat. NH₄Cl_((aq.)), H₂O,and brine. The organic phase was then dried over Na₂SO₄, which wassubsequently removed by vacuum filtration. The filtrate was concentratedin vacuo and the residue was dissolved in MeOH (1.5 mL). Compound 1 wasisolated from this solution by reverse-phase HPLC as a yellow solid (37mg, 37%). ¹H NMR (300 MHz, CD₃CN): d 8.50 (m, 1H), 8.11 (d, J=9.6 Hz,1H), 8.02 (s, 1H), 7.75 (s, 1H), 7.38 (s, 1H), 7.21 (dd, J=2.1, 9.3 Hz,1H), 7.00 (m, 1H), 6.03 (m, 1H), 5.97 (dt, J=6.9, 17.1 Hz, 1H), 5.67 (s,1H), 5.14 (d, J=17.1 Hz, 1H), 5.01 (d, J=11.4 Hz, 1H), 4.63 (m, 2H),4.44 (s, 1H), 4.17 (m, 2H), 4.08 (s, 1H), 4.04 (s, 3H), 2.74 (dd, J=7.2,14.1 Hz, 1H), 2.43 (ddd, J=3.3, 10.5, 13.8 Hz, 1H), 2.08 (m, 1H),1.24-1.75 (m, 19H), 1.15 (m, 1H), 1.04 (s, 9H). ³¹P NMR (121.4 MHz,CD₃CN) d 46.6. LC/MS=797 (M⁺+1), 819 (M⁺+Na)

Example 2 Preparation of Compound 2

Phosphonic acid intermediate III (208 mg, 0.64 mmol) was dissolved intoluene (8 mL). This solution was cooled to 0° C. and (COCl)₂ (111 μL,1.28 mmol) was added in a drop-wise fashion. DMF (22 μL, 0.28 mmol) wasthen added. The reaction was run for 2 h at 0° C. and determined to becomplete by ³¹P NMR.

³¹P NMR (121.4 MHz, CDCl₃): d 39.0, 38.5, 37.4, 36.5, 17.0, 16.2, 16.0,15.4.

The reaction was concentrated to orange-yellow oil and then placed underhigh vacuum for 1 h. The resulting residue was dissolved in THF (6.4 mL)and this solution was cooled to −78° C. A solution of EtLi in dibutylether (1.7 M, 566 μL, 0.96 mmol) was added drop-wise. After 40 min, moreEtLi (189 μL, 0.32 mmol) was added drop-wise. After 10 min, the reactionwas quenched at −78° C. by the addition of sat. NH₄Cl_((aq.)). Theorganic phase was diluted with EtOAc and extracted with sat.NH₄Cl_((aq.)) and brine. The organic phase was dried over MgSO₄.Concentration of the filtrate, after vacuum filtration removal of MgSO₄,yielded an orange oil from which the desired product was isolated bycolumn chromatography (SiO₂, 100% EtOAc) as a clear oil (67 mg, 31% over2 steps). ¹H NMR (300 MHz, CDCl₃) d 7.33 (s, 5H), 6.09 (dt, J=9.9, 17.1Hz, 1H diastereomer 1), 5.94 (dt, J=9.9, 17.1 Hz, 1H diastereomer 2),5.65 (d, J=23.7 Hz, 1H), 5.31 (d, J=17.1 Hz, 1H), 5.06 (m, 3H), 4.06 (m,2H), 2.09 (m, 1H), 1.73 (m, 2H), 1.50 (m, 2H), 1.25 (m, 4H), 1.13 (dt,J=8.1, 26.4 Hz, 3H). ³¹P NMR (121.4 MHz, CDCl₃) d 54.0, 53.6, 51.3, 50.8LC/MS=338 (M⁺+1), 360 (M⁺+Na)

A solution of(1-benzyloxycarbonylamino-2-vinyl-cyclopropyl)-ethyl-phosphinic acidethyl ester (104 mg, 0.308 mmol) in CH₃CN (7.7 mL) was cooled to 0° C.and TMSI (220 μL, 1.54 mmol) was added in a drop-wise fashion. Thereaction was warmed to rt and stirred for an hour. The reaction was thencooled to 0° C. and additional TMSI (110 μL, 0.77 mmol) was added in adrop-wise fashion. The reaction was warmed to rt and stirred for 30 min.The reaction was cooled back to 0° C. and 2,6-lutidine (360 μL, 3.1mmol) was added in a drop-wise fashion. This was followed by theaddition of Et₃N (1 mL, 7.2 mmol) and MeOH (4 mL). The reaction was thenconcentrated in vacuo and the crude amino phosphinic acid, which wasused directly in the next reaction.

A solution of dipeptide VII (81 mg, 0.123 mmol) in THF (2 mL) was cooledto −30° C. Et₃N (34 μL, 0.246 mmol) was added to this solution followedby ClCO₂Et (18 μL, 0.185 mmol). The reaction was stirred at atemperature between −20° C. and −30° C. for 30 min. Additional Et₃N (34μL, 0.246 mmol) and ClCO₂Et (18 μL, 0.185 mmol) was added to thereaction. The reaction was stirred for an additional 30 min at atemperature between −20° C. and −30° C. A solution of the crude aminophosphinic acid in CH₂Cl₂ (2 mL) was added in a drop-wise fashion at−30° C. and the reaction was warmed to rt. The reaction was quenched bythe addition of sat. NH₄Cl_((aq.)). The organic phase was diluted withEtOAc and washed with sat. NH₄Cl_((aq.)), H₂O, and brine. The organicphase was then dried over Na₂SO₄, which was subsequently removed byvacuum filtration. The filtrate was concentrated in vacuo and theresidue was dissolved in MeOH (1.5 mL). Compound 2 was isolated fromthis solution by reverse-phase HPLC as a yellow solid (37 mg, 37%). ¹HNMR (300 MHz, CDCl₃): d 8.27 (d, J=9.6 Hz, 1H), 8.18 (s, 1H), 7.75 (d,J=2.1 Hz, 1H), 7.73 (d, J=3.9 Hz, 1H), 7.31 (dd, J=2.1, 9.3 Hz, 1H),5.97 (dt, J=6.9, 17.1 Hz, 1H), 5.77 (s, 1H), 5.26 (d, J=17.1 Hz, 1H),5.08 (d, J=11.4 Hz, 1H), 4.63 (m, 2H), 4.44 (s, 1H), 4.17 (m, 2H), 4.08(s, 1H), 4.04 (s, 3H), 2.74 (dd, J=7.2, 14.1 Hz, 1H), 2.43 (ddd, J=3.3,10.5, 13.8 Hz, 1H), 2.08 (m, 1H), 1.84 (m, 2H), 1.54 (m, 8H), 1.34 (d,J=6.3 Hz, 6H), 1.34 (m, 2H), 1.15 (dt, J=7.8, 18.3 Hz, 3H), 1.04 (s,9H). ³¹P NMR (121.4 MHz, CDCl₃): d 50.6. LC/MS=811 (M⁺+1), 834 (M⁺+Na)

Example 3 Preparation of Compound 3

Phosphonic acid intermediate III (386 mg, 1.19 mmol) was dissolved intoluene (14.9 mL). This solution was cooled to 0° C. and (COCl)₂ (155μL, 1.78 mmol) was added in a drop-wise fashion. DMF (20 μL, 0.26 mmol)was then added. The reaction was run for 2 h at 0° C. and determined tobe complete by ³¹P NMR. ³¹P NMR (121.4 MHz, CDCl₃): d 39.0, 38.5, 37.4,36.6, 17.0, 16.2, 16.1, 15.4.

The reaction was concentrated to a yellow-orange oil and then placedunder high vacuum for 1 h. The resulting residue was dissolved in THF(11.9 mL) and this solution was cooled to −78° C. A 2.0 M solution ofn-BuLi in pentane (595 μL, 1.19 mmol) was added drop-wise. After 40 minmore n-BuLi (520 μL, 1.04 mmol) was added drop-wise. After 10 min thereaction was quenched at −78° C. by the addition of sat. NH₄Cl_((aq.)).The organic phase was diluted with EtOAc and extracted with sat.NH₄Cl_((aq.)) and brine. The organic phase was dried over MgSO₄.Concentration of the filtrate after vacuum filtration removal of theMgSO₄ yielded an orange oil from which the product was isolated bycolumn chromatography (SiO₂, 7/3 EtOAc:hexane) as a clear oil (243 mg,56% over 2 steps). ¹H NMR (300 MHz, CDCl₃): d 7.35 (s, 5H), 6.12 (dt,J=9.9, 16.8 Hz, 1H diastereomer 1), 5.96 (dt, J=10.2, 16.8 Hz, 1Hdiastereomer 2), 5.33 (m, 2H), 5.09 (m, 3H), 4.11 (m, 2H), 2.01 (brd,J=6.6 Hz, 1H), 1.50-1.90 (m, 6H), 1.37 (brd, J=5.1 Hz, 2H), 1.26(quart., J=6.2 Hz, 3H), 0.9 (m, 3H)

³¹P NMR (121.4 MHz, CDCl₃): d 52.8, 52.4, 50.2, 49.7

LC/MS=366 (M⁺+1), 388 (M⁺+Na)

A solution of(1-benzyloxycarbonylamino-2-vinyl-cyclopropyl)-butyl-phosphinic acidethyl ester (364 mg, 0.996 mmol) in CH₃CN (25 mL) was cooled to 0° C.and TMSI (220 μL, 1.54 mmol) was added in a drop-wise fashion. Thereaction was warmed to rt and stirred for an hour. The reaction was thencooled to 0° C. and additional TMSI (711 μL, 4.98 mmol) was added in adrop-wise fashion. The reaction was warmed to rt and stirred for 1 h.The reaction was cooled back to 0° C. and 2,6-lutidine (1 mL, 10.1 mmol)was added in a drop-wise fashion. This was followed by the addition ofEt₃N (1 mL, 7.2 mmol) and MeOH (4 mL). The reaction was warmed to rt andthen concentrated in vacuo. The crude mixture was used directly in thenext reaction.

A solution of the starting dipeptide VII (100 mg, 0.153 mmol) in THF (2mL) was cooled to −30° C. Et₃N (32 μL, 0.230 mmol) was added to thissolution followed by ClCO₂Et (22 μL, 0.23 mmol). The reaction wasstirred at a temperature between −20° C. and −30° C. for 30 min.Additional Et₃N (32 μL, 0.23 mmol) and ClCO₂Et (22 μL, 0.23 mmol) wasadded to the reaction. The reaction was stirred for an additional 30 minat a temperature between −20° C. and −30° C. A solution of crude productfrom step 1 in CH₂Cl₂ (2 mL) was added in a drop-wise fashion at −30° C.and the reaction was warmed to rt. The reaction was quenched by theaddition of sat. NH₄Cl_((aq.)). The organic phase was diluted with EtOAcand extracted with sat. NH₄Cl_((aq.)), H₂O, and brine. The organic phasewas then dried over Na₂SO₄, which was subsequently removed by vacuumfiltration. The filtrate was concentrated in vacuo and the residue wasdissolved in MeOH (1.5 mL). The desired product from the coupling wasisolated by reverse-phase HPLC. This coupling reaction was repeated oncemore on the same scale and the isolated mixture of products from bothreaction runs were combined.

The combined products from the coupling reactions were dissolved inCH₃CN (5.4 mL) and 2,6-lutidine (149 μL, 1.29 mmol) was then added. Thissolution was cooled to 0° C. and TMSI (184 μL, 1.29 mmol) was added in adrop-wise fashion. The reaction was stirred at rt for 1 h and thencooled to 0° C. Additional 2,6-lutidine (125 μL, 0.645 mmol) and TMSI(92 μL, 0.645 mmol) was added and the reaction was warmed to rt. Thereaction was then cooled to 0° C. and Et₃N (1.5 mL, 20.4 mmol) was addedin a drop-wise fashion followed by MeOH (5 mL). The reaction wasevaporated in vacuo and then dissolved in MeOH (1.5 mL). Compound 3 wasisolated from this solution by reverse-phase HPLC as a yellow solid (86mg, 33% over 2 steps). ¹H NMR (300 MHz, CDCl₃): d 8.26 (d, J=9 Hz, 1H),8.15 (s, 1H), 7.70 (d, J=2.1 Hz, 2H), 7.24 (dd, J=2.1, 9 Hz, 1H), 5.93(dt, J=9.6, 19.5 Hz, 1H), 5.71 (s, 1H), 5.11 (d, J=16.8 Hz, 1H), 4.95(d, J=12.3 Hz, 1H), 4.70 (d, J=12.3 Hz, 1H), 4.62 (dd, J=7.2, 9.3 Hz,1H), 4.51 (s, 1H), 4.21 (s, 1H), 4.14 (q, J=6.6 Hz, 1H), 4.07 (dd,J=2.4, 9.9 Hz, 1H), 4.02 (s, 3H), 2.82 (dd, J=7.5, 14.4 Hz, 1H), 2.45(ddd, J=3.9, 10.2, 14.1 Hz, 1H), 1.98 (m, 1H), 1.40-1.80 (m, 13H), 1.34(d, J=6.3 Hz, 6H), 1.14-1.32 (m, 3H), 1.01 (s, 9H), 0.86 (t, J=7.2 Hz,3H). ³¹P NMR (121.4 MHz, CDCl₃): d 43.1. LC/MS=839 (M⁺+1), 861 (M⁺+Na).

Example 4 Preparation of Compound 4

Phosphonic acid intermediate III (415 mg, 1.28 mmol) was dissolved intoluene (8 mL). This solution was cooled to 0° C. and (COCl)₂ (222 μL,2.56 mmol) was added in a drop-wise fashion. DMF (44 μL, 0.56 mmol) wasthen added. The reaction was run for 2 h at 0° C. and determined to becomplete by ³¹P NMR. ³¹P NMR (121.4 MHz, CDCl₃): d 39.0, 38.5, 37.4,36.5, 17.0, 16.2, 16.0, 15.4.

The reaction was concentrated to an orange-yellow oil and then placedunder high vacuum for 1 h. The resulting residue was dissolved in THF(6.4 mL) and this solution was cooled to −78° C. A 1.4 M solution ofsec-butyllithium in cyclohexane (1.37 mL, 1.92 mmol) was addeddrop-wise. After 40 min more sec-butyllithium in cyclohexane (456 μL,0.64 mmol) was added drop-wise. After 10 min the reaction was quenchedat −78° C. by the addition of sat. NH₄Cl_((aq.)). The organic phase wasdiluted with EtOAc and extracted with sat. NH₄Cl_((aq.)) and brine. Theorganic phase was dried over MgSO₄. Concentration of the filtrate aftervacuum filtration removal of the MgSO₄ yielded an orange oil from whichthe product was isolated by column chromatography (SiO₂, 60% EtOAc inHexane) as a clear oil (146 mg, 31% over 2 steps).

¹H NMR (300 MHz, CDCl₃): d 7.33 (s, 5H), 6.07 (dt, J=9.9, 17.1 Hz, 1H),5.55 (d, J=23.7 Hz, 1H), 5.31 (d, J=17.1 Hz, 1H), 5.06 (m, 3H), 4.06 (m,2H), 2.09 (m, 1H), 1.65-1.83 (m, 3H), 1.58 (m, 1H) 1.41 (m, 1H),1.03-1.32 (m, 6H), 0.97 (dt, J=8.1, 26.4 Hz, 3H). ³¹P NMR (121.4 MHz,CDCl₃): d 54.9, 54.3, 50.8, 50.0

LC/MS=366 (M⁺+1), 388 (M⁺+Na)

A solution of(1-benzyloxycarbonylamino-2-vinyl-cyclopropyl)-sec-butyl-phosphinic acidethyl ester (112 mg, 0.308 mmol) in CH₃CN (7.7 mL) was cooled to 0° C.and TMSI (220 μL, 1.54 mmol) was added in a drop-wise fashion. Thereaction was warmed to rt and stirred for an hour. The reaction was thencooled to 0° C. and additional TMSI (110 μL, 0.77 mmol) was added in adrop-wise fashion. The reaction was warmed to rt and stirred for 30 min.The reaction was cooled back to 0° C. and 2,6-lutidine (360 μL, 3.1mmol) was added in a drop-wise fashion. This was followed by theaddition of Et₃N (1 mL, 7.2 mmol) and MeOH (4 mL). The reaction was thenconcentrated in vacuo and the resulting crude product was used directlyin the next reaction.

A solution of dipeptide VII (81 mg, 0.123 mmol) in THF (2 mL) was cooledto −30° C. Et₃N (34 μL, 0.246 mmol) was added to this solution followedby ClCO₂Et (18 μL, 0.185 mmol). The reaction was stirred at atemperature between −20° C. and −30° C. for 30 min. Additional Et₃N (34μL, 0.246 mmol) and ClCO₂Et (18 μL, 0.185 mmol) was added to thereaction. The reaction was stirred for an additional 30 min at atemperature between −20° C. and −30° C. A solution of crude product fromstep 1 in CH₂Cl₂ (2 mL) was added in a drop-wise fashion at −30° C. andthe reaction was warmed to rt and stirred for 2 hours. The reaction wasquenched by the addition of sat. NH₄Cl_((aq.)). The organic phase wasdiluted with EtOAc and extracted with sat. NH₄Cl_((aq.)), H₂O, andbrine. The organic phase was dried over Na₂SO₄, which was subsequentlyremoved by vacuum filtration. The filtrate was concentrated in vacuo andthe residue was dissolved in MeOH (1.5 mL). Compound 4 was isolated fromthis solution by reverse-phase HPLC as a yellow solid (42 mg, 41%). ¹HNMR (300 MHz, CD₃OD): d 8.27 (d, J=9.6 Hz, 1H), 8.18 (s, 1H), 7.75 (d,J=2.1 Hz, 1H), 7.39 (d, J=3.9 Hz, 1H), 7.31 (dd, J=2.1, 9.3 Hz, 1H),6.01 (dt, J=6.9, 17.1 Hz, 1H), 5.77 (s, 1H), 5.26 (d, J=17.1 Hz, 1H),5.08 (d, J=11.4 Hz, 1H), 4.63 (m, 2H), 4.44 (s, 1H), 4.17 (m, 2H), 4.08(s, 1H), 4.04 (s, 3H), 2.76 (dd, J=7.2, 14.1 Hz, 1H), 2.43 (ddd, J=3.3,10.5, 13.8 Hz, 1H), 2.08 (m, 1H), 1.96 (m, 2H), 1.60-1.82 (m, 9H), 1.34(d, J=6.3 Hz, 6H), 1.22 (m, 6H), 1.04 (s, 9H), 0.99 (m, 3H). ³¹P NMR(121.4 MHz, CD₃OD) d 52.4, 52.2

LC/MS=839 (M⁺+1), 861 (M⁺+Na)

Example 5 Preparation of Compound 5

Phosphonic acid intermediate III (415 mg, 1.28 mmol) was dissolved intoluene (8 mL). This solution was cooled to 0° C. and (COCl)₂ (222 μL,2.56 mmol) was added in a drop-wise fashion. DMF (44 μL, 0.56 mmol) wasthen added. The reaction was run for 2 h at 0° C. and determined to becomplete by ³¹P NMR.

³¹P NMR (121.4 MHz, CDCl₃): d 39.0, 38.5, 37.4, 36.5, 17.0, 16.2, 16.0,15.4.

The reaction was concentrated to an orange-yellow oil and then placedunder high vacuum for 1 h. The resulting residue was dissolved in THF(6.4 mL) and this solution was cooled to −78° C. A 0.7 M solution ofisopropyllithium in pentane (2.74 mL, 1.92 mmol) was added drop-wise.After 40 min more isopropyllithium (912 μL, 0.64 mmol) was addeddrop-wise. After 10 min the reaction was quenched at −78° C. by theaddition of sat. NH₄Cl_((aq.)). The organic phase was diluted with EtOAcand extracted with sat. NH₄Cl_((aq.)) and brine. The organic phase wasdried over MgSO₄. Concentration of the filtrate after vacuum filtrationremoval of the MgSO₄ yielded an orange oil from which the product wasisolated by column chromatography (SiO₂, 100% EtOAc) as a clear oil (200mg, 45% over 2 steps). ¹H NMR (300 MHz, CD₃CN): d 7.38 (s, 5H), 6.69 (m,1H), 6.12 (m, 1H), 5.35 (m, 1H), 5.06 (m, 4H), 4.06 (m, 2H), 2.09 (m,1H), 1.55 (m, 1H) 1.41 (m, 1H), 1.02-1.35 (m, 9H). ³¹P NMR (121.4 MHz,CD₃CN): d 56.0, 53.8. LC/MS=352 (M⁺+1), 374 (M⁺+Na)

A solution of(1-benzyloxycarbonylamino-2-vinyl-cyclopropyl)-isopropyl-phosphinic acidethyl ester (108 mg, 0.308 mmol) in CH₃CN (7.7 mL) was cooled to 0° C.and TMSI (220 μL, 1.54 mmol) was added in a drop-wise fashion. Thereaction was warmed to rt and stirred for an hour. The reaction was thencooled to 0° C. and additional TMSI (110 μL, 0.77 mmol) was added in adrop-wise fashion. The reaction was warmed to rt and stirred for 30 min.The reaction was cooled back to 0° C. and 2,6-lutidine (360 μL, 3.1mmol) was added in a drop-wise fashion. This was followed by theaddition of Et₃N (1 mL, 7.2 mmol) and MeOH (4 mL). The reaction was thenconcentrated in vacuo and the crude product was used directly in thenext reaction.

A solution of VII (81 mg, 0.123 mmol) in THF (2 mL) was cooled to −30°C. Et₃N (34 μL, 0.246 mmol) was added to this solution followed byClCO₂Et (18 μL, 0.185 mmol). The reaction was stirred at a temperaturebetween −20° C. and −30° C. for 30 min. Additional Et₃N (34 μL, 0.246mmol) and ClCO₂Et (18 μL, 0.185 mmol) was added to the reaction. Thereaction was stirred for an additional 30 min at a temperature between−20° C. and −30° C. A solution of crude product from step 1 in CH₂Cl₂ (2mL) was added in a drop-wise fashion at −30° C. and the reaction waswarmed to rt and stirred for 2 hours. The reaction was quenched by theaddition of sat. NH₄Cl_((aq.)). The organic phase was diluted with EtOAcand extracted with sat. NH₄Cl_((aq.)), H₂O, and brine. The organic phasewas then dried over Na₂SO₄, which was subsequently removed by vacuumfiltration. The filtrate was concentrated in vacuo and the residue wasdissolved in MeOH (1.5 mL). Compound 5 was isolated from this solutionby reverse-phase HPLC as a yellow solid (40 mg, 40%). ¹H NMR (300 MHz,CD₃CN): d 8.27 (d, J=9.6 Hz, 1H), 8.11 (m, 1H), 8.05 (s, 1H), 7.75 (d,J=2.1 Hz, 1H), 7.53 (d, J=3.9 Hz, 1H), 7.³¹ (dd, J=2.1, 9.3 Hz, 1H),6.75 (m, 1H), 6.06 (dt, J=6.9, 17.1 Hz, 1H), 5.77 (m, 2H), 5.26 (d,J=17.1 Hz, 1H), 5.08 (d, J=11.4 Hz, 1H), 4.63 (m, 2H), 4.17 (m, 2H),4.08 (s, 1H), 4.04 (s, 3H), 2.74 (dd, J=7.2, 14.1 Hz, 1H), 2.53 (ddd,J=3.3, 10.5, 13.8 Hz, 1H), 2.21 (m, 1H), 2.08 (m, 1H), 1.42-1.78 (m,8H), 1.34 (d, J=6.3 Hz, 6H), 1.34 (m, 2H) 1.15 (m, 5H), 1.04 (s, 9H),0.99-1.03 (m, 3H). ³¹P NMR (121.4 MHz, CD₃CN): d 50.6. LC/MS=825 (M⁺+1),847 (M⁺+Na)

Example 6 Preparation of Compound 6

Phosphonic acid intermediate III (415 mg, 1.28 mmol) was dissolved intoluene (8 mL). This solution was cooled to 0° C. and (COCl)₂ (222 μL,2.56 mmol) was added in a drop-wise fashion. DMF (44 μL, 0.56 mmol) wasthen added. The reaction was run for 2 h at 0° C. and determined to becomplete by ³¹P NMR.

³¹P NMR (121.4 MHz, CDCl₃): d 39.0, 38.5, 37.4, 36.5, 17.0, 16.2, 16.0,15.4.

The reaction was concentrated to an orange-yellow oil and then placedunder high vacuum for 1 h. The resulting residue was dissolved in THF(6.4 mL) and this solution was cooled to −78° C. A 1.0 M solution ofvinylmagnesium bromide in tetrahydrofuran (2.6 mL, 2.6 mmol) was addeddrop-wise. After 40 min more vinylmagnesium bromide (2.6 mL, 2.6 mmol)was added drop-wise. After 10 min the reaction was quenched at −78° C.by the addition of sat. NH₄Cl_((aq.)). The organic phase was dilutedwith EtOAc and extracted with sat. NH₄Cl_((aq.)) and brine. The organicphase was dried over MgSO₄. Concentration of the filtrate after vacuumfiltration removal of the MgSO₄ yielded an orange oil from which theproduct was isolated by column chromatography (SiO₂, 100% EtOAc) as aclear oil (214 mg, 40% over 2 steps). ¹H NMR (300 MHz, CDCl₃): d 7.33(s, 5H), 6.09-6.15 (m, 2H), 5.55 (m, 1H), 5.31 (m, 1H), 5.05 (m, 4H),4.06 (m, 2H), 2.09 (m, 1H), 1.73 (m, 1H), 1.60 (m, 1H), 1.43 (m, 1H),1.13 (dt, J=8.1, 26, 4 Hz, 3H)

³¹P NMR (121.4 MHz, CDCl₃): d 36.5, 34.6. LC/MS=336 (M⁺+1), 358 (M⁺+Na).

A solution of(1-benzyloxycarbonylamino-2-vinyl-cyclopropyl)-vinyl-phosphinic acidethyl ester (103 mg, 0.308 mmol) in CH₃CN (7.7 mL) was cooled to 0° C.and TMSI (220 μL, 1.54 mmol) was added in a drop-wise fashion. Thereaction was warmed to rt and stirred for an hour. The reaction was thencooled to 0° C. and additional TMSI (110 μL, 0.77 mmol) was added in adrop-wise fashion. The reaction was warmed to rt and stirred for 30 min.The reaction was cooled back to 0° C. and 2,6-lutidine (360 μL, 3.1mmol) was added in a drop-wise fashion. This was followed by theaddition of Et₃N (1 mL, 7.2 mmol) and MeOH (4 mL). The reaction was thenconcentrated in vacuo and crude was used directly in the next reaction.

A solution of dipeptide VII (81 mg, 0.123 mmol) in THF (2 mL) was cooledto −30° C. Et₃N (34 μL, 0.246 mmol) was added to this solution followedby ClCO₂Et (18 μL, 0.185 mmol). The reaction was stirred at atemperature between −20° C. and −30° C. for 30 min. Additional Et₃N (34μL, 0.246 mmol) and ClCO₂Et (18 μL, 0.185 mmol) was added to thereaction. The reaction was stirred for an additional 30 min at atemperature between −20° C. and −30° C. A solution of crude from step 1in CH₂Cl₂ (2 mL) was added in a drop-wise fashion at −30° C. and thereaction was warmed to rt and stirred for 2 hours. The reaction wasquenched by the addition of sat. NH₄Cl_((aq.)). The organic phase wasdiluted with EtOAc and extracted with sat. NH₄Cl_((aq.)), H₂O, andbrine. The organic phase was then dried over Na₂SO₄, which wassubsequently removed by vacuum filtration. The filtrate was concentratedin vacuo and the residue was dissolved in MeOH (1.5 mL). Compound 6 wasisolated from this solution by reverse-phase HPLC as a yellow solid (45mg, 45%). ¹H NMR (300 MHz, CD₃CN) d 8.25 (br, 1H), 8.20 (d, J=9.6 Hz,1H), 8.02 (s, 1H), 7.75 (s, 1H), 7.39 (s, 1H), 7.23 (dd, J=2.1, 9.3 Hz,1H), 6.84 (br, 1H), 6.35 (m, 2H), 5.97 (m, 3H), 5.77 (m, 1H), 5.61 (s,1H), 5.26 (d, J=17.1 Hz, 1H), 5.08 (d, J=11.4 Hz, 1H), 4.63 (m, 2H),4.44 (s, 1H), 4.17 (m, 2H), 4.08 (s, 1H), 4.04 (s, 3H), 2.74 (dd, J=7.2,14.1 Hz, 1H), 2.43 (ddd, J=3.3, 10.5, 13.8 Hz, 1H), 1.41-1.78 (m, 8H),1.34 (d, J=6.3 Hz, 6H), 1.34 (m, 2H), 1.15 (m, 1H), 1.04 (s, 9H)

³¹P NMR (121.4 MHz, CD₃CN) d 30.2. LC/MS=809 (M⁺+1), 831 (M⁺+Na).

Example 7 Preparation of Compound 7

Phosphonic acid intermediate III (451 mg, 1.39 mmol) was dissolved intoluene (17.4 mL). This solution was cooled to 0° C. and (COCl)₂ (1.21mL, 13.87 mmol) was added in a drop-wise fashion. DMF (24 μL, 0.306mmol) was then added. The reaction was run for 2 h at 0° C. and then 18h at rt. The reaction was determined to be complete by ³¹P NMR. ³¹P NMR(121.4 MHz, CDCl₃) d 39.3, 38.8, 37.6, 36.8, 17.2, 16.4, 16.3, 15.6.

The reaction was concentrated to an orange-yellow oil and then placedunder high vacuum for 1 h. The resulting residue was dissolved in THF(13.9 mL) and this solution was cooled to −78° C. A 1.8M solution ofPhLi in Et₂O (1.2 mL, 2.17 mmol) was added drop-wise. After 30 min thereaction was quenched at −78° C. by the addition of sat. NH₄Cl_((aq.)).The organic phase was diluted with EtOAc and extracted with sat.NH₄Cl_((aq.)) and brine. The organic phase was dried over MgSO₄ whichwas subsequently removed by vacuum filtration. Concentration of thefiltrate yielded an orange oil from which the desired product wasisolated by column chromatography (SiO₂, 7/3 EtOAc:hexane) as a clearoil (243 mg, 56% over 2 steps) in 73% purity by ³¹P NMR. ¹H NMR (300MHz, CDCl₃) d 7.75 (m, 2H), 7.56 (m, 1H), 7.20-7.44 (m, 7H), 6.18 (m,1H), 5.39 (d, J=17.1 Hz, 1H), 4.80-5.30 (m, 4H), 4.0-4.3 (m, 2H), 1.91(m, 1H), 1.69 (m, 1H), 1.2-1.4 (m, 4H). ³¹P NMR (121.4 MHz, CDCl₃) d37.8, 37.4, 36.2, 36.0, 35.0, 34.7, 33.4, 33.3

LC/MS=386 (M⁺+1), 408 (M⁺+Na).

A solution of(1-benzyloxycarbonylamino-2-vinyl-cyclopropyl)-phenyl-phosphinic acidethyl ester (150 mg, 0.389 mmol) in ACN (10 mL) was cooled to 0° C. andTMSI (278 μL, 1.95 mmol) was added in a drop-wise fashion. The reactionwas warmed to rt and stirred for an hour. The reaction was cooled backto 0° C. and Et₃N (1.5 mL, 20.4 mmol) and MeOH (5 mL) were added in adrop-wise fashion. The reaction was then concentrated in vacuo and thecrude product was used directly in the next reaction.

A solution of dipeptide VII (50 mg, 0.076 mmol) in THF (2 mL) was cooledto −30° C. Et₃N (16 μL, 0.114 mmol) was added to this solution followedby ClCO₂Et (15 μL, 0.114 mmol). The reaction was stirred at atemperature between −20° C. and −30° C. for 30 min. Additional Et₃N (16μL, 0.114 mmol) and ClCO₂Et (15 μL, 0.114 mmol) was added to thereaction. The reaction was stirred for an additional 30 min at atemperature between −20° C. and −30° C. A solution of the crude productfrom step 1 in CH₂Cl₂ (2 mL) was added in a drop-wise fashion at −30° C.and the reaction was warmed to rt. The reaction was quenched by theaddition of sat. NH₄Cl_((aq.)). The organic phase was diluted with EtOAcand extracted with sat. NH₄Cl_((aq.)), H₂O, and brine. The organic phasewas then dried over Na₂SO₄, which was subsequently removed by vacuumfiltration. The filtrate was concentrated in vacuo and the residue wasdissolved in MeOH (1.5 mL). Compound 7 was isolated from this solutionby reverse-phase HPLC as a yellow solid (17 mg, 25%). ¹H NMR (300 MHz,CDCl₃) d 8.22 (d, J=9.6 Hz, 1H), 8.18 (s, 1H), 7.89 (dd, J=6.9, 11.7 Hz,2H), 7.74 (d, J=2.1 Hz, 1H), 7.72 (s, 1H), 7.53 (m, 3H), 7.30 (dd,J=2.1, 9 Hz, 1H), 6.14 (dt, J=10.2, 19.5 Hz, 1H), 5.71 (s, 1H), 5.22 (d,J=17.1 Hz, 1H), 5.02 (d, J=10.2 Hz, 1H), 4.55 (m, 2H), 4.40 (s, 1H),4.18 (quint., J=6.6 Hz, 1H), 4.11 (s, 1H), 4.04 (m, 4H), 5.60 (dd,J=6.9, 14.1 Hz, 1H), 2.23 (ddd, J=3.6, 10.2, 13.8 Hz, 1H), 2.12 (m, 1H),1.72 (m, 1H), 1.40-1.66 (m, 9H), 1.34 (d, J=6.3 Hz, 6H), 1.03 (s, 9H).³¹P NMR (121.4 MHz, CDCl₃) d 34.0

LC/MS=859 (M⁺+1), 881 (M⁺+Na).

Example 8 Preparation of Compound 8

To a solution of 1 (677 mg, 0.79 mmol) in DME (5 mL) and H₂O (0.4 mL)was added p-tosylhydrazide (737 mg, 3.96 mmol) and NaOAc (650 mg, 7.93mmol). The reaction mixture was heated to 95° C. for 1.5 h and cooled tort. A few drops of 3 N HCl was added to adjust the pH to 2. The crudeproduct was purified by HPLC to give 8 (587 mg, 76%) as a yellow solid.¹H NMR (300 MHz, CD₃CN): d 8.16 (m, 1H), 8.06 (s, 1H), 7.71 (s, 1H),7.44 (s, 1H), 7.25 (m, 1H), 6.83 (m, 1H), 5.92 (m, 1H), 5.61 (br, 1H),4.58 (m, 2H), 4.41 (m, 1H), 4.14 (m, 2H), 4.05 (m, 4H), 2.76 (m, 1H),2.45 (m, 1H), 1.80 (m, 2H), 1.65 (m, 10H) 1.38 (d, J=6.3, 6H), 1.21 (m,1H), 0.98 (m, 12H). ³¹P NMR (121.4 MHz, CD₃CN):

46.569. LC/MS=799 (M⁺+1)

Example 9 Preparation of Compound 9

A solution of phosphonous acid (IV) (150 mg, 0.48 mmol) in CH₃CN (1 mL)was stirred at 0° C. as iodotrimethylsilane (345 μl, 2.42 mmol) wasadded. The solution stirred at rt for 45 min. and was cooled again to 0°C. and triethylamine (1 mL, 7.33 mmol) and MeOH (2 mL) was added. Thesolution was warmed to rt and stirred for an additional 20 minutes. Thesolution was then concentrated, azeotroped with toluene (×2) and driedon high vacuum for 30 minutes. The crude was coupled with VII (209 mg,0.32 mmol), using HATU (304 mg, 0.80 mmol), and NMM (176 μl, 1.60 mmol)in DMF (1 mL) overnight at rt. The reaction was concentrated andpurified with a Gilson HPLC to obtain 9 as a yellow solid. ¹H NMR (300MHz, CD₃OD): d 8.18 (s, 1H), 8.20 (m, 2H), 7.78 (s, 1H), 7.38 (m, 1H),6.20 (s, J=9.2 Hz, 1H), 5.90 (m, 1H), 5.80 (bs, 1H), 5.23 (dd, 1H), 5.18(d, J=9.0 Hz, 1H) 4.78 (s, 1H), 4.58 (m, 1H), 4.30 (m, 1H), 4.20 (q,2H), 4.05 (m, 2H), 4.01 (s, 3H), 2.79 (m, 1H), 2.57 (m, 1H), 2.15 (m,1H), 1.62 (m, 2H), 1.50 (m, 2H) 1.30 (d, 3H), 1.05 (s, 9H). ³¹P (121.4MHz, CD₃OD):

22.768, 22.682

Example 10 Preparation of Compound 10

Same procedures as described for example 6 was used to provide compound10. ¹H NMR (300 MHz, CD₃OD)

8.25 (d J=9 Hz, 1H), 8.18 (s, 1H), 7.75 (m, 2H), 7.30 (dd, J=9.3, 2.1Hz, 1H), 5.96 (dt, J=6.9, 19.8 Hz, 1H), 5.77 (s, 1H), 5.25 (d, J=17.1Hz, 1H), 5.06 (d, J=10.5 Hz, 1H), 4.64 (m, 2H), 4.44 (s, 1H), 4.15 (m,3H), 4.04 (m, 3H), 2.76 (dd, J=7.5, 14.1 Hz, 1H), 2.43 (ddd, J=3.9,10.2, 13.8 Hz, 1H), 2.08 (m, 1H), 1.4-1.9 (brm, 14H), 1.34 (d, J=6.3 Hz,6H), 1.04 (s, 9H), 1.02 (m, 3H).

³¹P NMR (121.4 MHz, CD₃OD) d 48.8

LC/MS=825.3 (M⁺+1), 847.2 (M⁺+Na)

Example 11 Preparation of Compound 11

A solution of the phosphonous acid IV (499 mg, 1.61 mmol), Hunig's Base(794 μL, 3.88 mmol), and chlorotrimethylsilane (590 μL, 3.57 mmol) inCH₂Cl₂ (7.5 mL) was stirred at r.t. for 30 minutes before ethyl2-bromoacetate (395 μL, 3.65 mmol) was added. The solution was heated at40° C. for 7.5 h. The solution was concentrated and the residue wasdissolved in ethyl acetate (30 mL) and then washed with H₂O (30 mL×2).The aqueous layers were extracted with ethyl acetate (30 mL). Thecombined organic layers were dried (MgSO₄) and concentrated. The residuewas purified by column chromatography using hexane:ethyl acetate aseluent to obtain[(1-benzyloxycarbonylamino-2-vinyl-cyclopropyl)-ethoxy-phosphinoyl]-aceticacid ethyl ester (344 mg, 54%). ¹H NMR (300 MHz, CDCl₃): d 7.34 (s, 5H),6.04 (m, 0.39H), 5.91 (m, 0.53H), 5.72 (d, 1H), 5.42 (s, 1H), 5.36 (s,1H), 5.30 (s, 2H), 5.09 (m, 3H), 4.18 (m, 4H), 3.04 (m, 2H), 2.30 (m,1H), 2.03 (m, 1H), 1.85 (m, 2H), 1.70 (m, 2H), 1.58 (m, 2H), 1.38 (m,2H), 1.25 (m, 6H). ³¹P (121.4 MHz, CDCl₃):

43.406, 42.646, 39.087

A solution of[(1-benzyloxycarbonylamino-2-vinyl-cyclopropyl)-ethoxy-phosphinoyl]-aceticacid ethyl ester (352 mg, 0.89 mmol) in THF (3 mL) was stirred at 0° C.as 1 N NaOH (980 μL, 0.98 mmol) was added. The solution was stirredovernight at r.t. and then concentrated, diluted with H₂O (10 mL) andwashed with ethyl acetate. The aqueous layer was acidified with 1 N HCl(5 mL) and extracted with ethyl acetate (×2). The organic extracts werewashed with H₂O, dried (MgSO₄) and concentrated to yield[(1-Benzyloxycarbonylamino-2-vinyl-cyclopropyl)-ethoxy-phosphinoyl]-aceticacid (224 mg, 69%). ¹H NMR (300 MHz, CDCl₃): d 7.34 (s, 5H), 5.91 (m,2H), 5.20 (m, 2H), 4.21 (m, 2H), 3.11 (m, 2H), 2.30 (m, 1H), 2.03 (m,1H), 1.85 (m, 2H), 1.70 (m, 2H), 1.58 (m, 2H), 1.38 (m, 2H), 1.25 (m,3H). ³¹P (121.4 MHz, CDCl₃):

45.109, 41.119, 40.965, 39.514

A solution of the acid (224 mg, 0.61 mmol), dimethylammonium chloride(125 mg, 1.53 mmol), HATU (697 mg, 1.83 mmol) and N-methylmorpholine(600 μl, 5.46 mmol) was stirred in DMF (3 mL) at rt for 2.5 h. Thesolution was concentrated and the residue was dissolved in ethyl acetate(30 mL) and washed with H₂O (2×30 mL) and brine. The aqueous layer wasextracted with ethyl acetate (30 mL) and the combined organic extractswere dried (MgSO₄) and concentrated. The residue was triturated withCH₂Cl₂ (10 mL) and filtered. The filtrate was concentrated and theresidue treated with CH₂Cl₂ and then filtered. The desired product (240mg, 99%) was isolated by column chromatography using hexane:ethylacetate as eluent.

¹H NMR (300 MHz, CDCl₃): δ 7.33 (s, 5H), 6.38 (s, 1H), 6.00 (m, 1H),5.44 (s, 1H), 5.38 (s, 1H), 5.30 (s, 2H), 5.04 (m, 4H), 4.23 (m, 2H),3.18 (s, 1.08H), 3.09 (s, 1.62H), 2.88 (s, 1.08H), 2.81 (s, 1.62H), 2.38(m, 1H), 1.87 (min, 1H), 1.76 (m, 1H), 1.45 (m, 1H), 1.23 (m, 3H). ³¹P(121.4 MHz, CDCl₃):

46.664, 45.538, 42.765, 42.417. Deprotection and coupling tointermediate VII gave 11.

LC/MS=868 (M⁺+1),

Example 12 Preparation of Compound 12

Intermediate IV (840 mg, 2.7 mmol) was dissolved in 8 mL of dry THF andcooled to −40° C. 1N NaHMDS (4.1 mL, 4.1 mmol) in THF was added dropwiseand the reaction was stirred at −40° C. for 30 minutes.1-bromo-2-methyl-propene (446 μl, 4.1 mmol) was dissolved in 1 mL of THFand then added dropwise. The reaction was allowed to warm to rt. Afterstirring at rt overnight, the reaction was cooled to 0° C. and 3001 ofacetic acid was added to quench the reaction. The mixture was thenconcentrated in vacuo and diluted with ethyl acetate. The organic wasthen extracted once with water and once with brine. The organic was thendried over Mg₂SO₄ and concentrated in vacuo to afford an oily residue.The product (83 mg, 9%) was then isolated from the residue by silica gelchromatography (3:1-ethyl acetate:hexane). ¹H NMR (300 MHz, CDCl₃) d7.36 (s, 3H), 6.0 (m, 1H), 5.30 (m, 2H), 5.10 (b, 4H), 4.13 (m, 2H),2.13 (m, 2H), 1.79 (m, 3H), 1.54-1.39 (m, 4H), 1.28 (m, 3H), 1.03 (m,6H). ³¹P NMR (121.4 MHz, CDCl₃) d 50.26, 47.54. LC/MS=366 (M⁺+1)

Coupling to dipeptide VII as described above gave compound 12 (28 mg,28%). ¹H NMR (300 MHz, CD₃OD) 8.27 (d, J=9.3 Hz, 1H), 8.22 (s, 1H), 7.76(s, 2H), 7.³¹ (d, J=9.0 Hz, 1H), 5.94 (m, 1H), 5.79 (b, 1H), 5.25 (d,J=17.1 Hz, 1H), 5.07 (d, J=9.9 Hz, 1H), 4.64 (m, 2H), 4.43 (s, 1H), 4.15(m, 2H), 4.11 (s, 1H), 4.04 (s, 1H), 2.80 (m, 2H), 2.45 (m, 1H), 2.15(m, 1H), 1.75 (m, 1H), 1.60-1.40 (m, 8H), 1.35 (d, J=6.3 Hz, 12H), 1.04(s, 10H). ³¹P NMR (121.4 MHz, CD₃OD) d 24.48. LC/MS=839 (M⁺+1)

Example 13 Preparation of Compound 13

Examples 13 through 15 were prepared by the same method as example 12.¹H NMR (300 MHz, CD₃OD): d 8.26 (m, 1H), 8.17 (m, 1H), 7.76 (s, 2H),7.32 (m, 1H), 5.95 (m, 1H), 5.80 (br, 1H), 5.36 (m, 1H), 5.13 (m, 1H),4.63 (m, 2H), 4.41 (m, 1H), 4.15 (m, 2H), 4.04 (m, 4H), 2.66 (m, 1H),2.33 (m, 1H), 1.94 (m, 2H), 1.65 (m, 13H) 1.38 (d, J=6.3 Hz, 6H), 1.04(s, 9H). ³¹P (121.4 MHz, CD₃OD):

33.642. LC/MS=837 (M⁺+1)

Example 14 Preparation of Compound 14

¹H NMR (300 MHz, CD₃OD): d 8.27 (d, J=9.6 Hz, 1H), 8.18 (m, 1H), 7.74(s, 2H), 7.32 (m, 1H), 5.89 (m, 2H), 5.78 (br, 1H), 5.26 (m, 1H), 5.09(m, 2H), 4.97 (m, 1H), 4.65 (m, 2H), 4.44 (m, 1H), 4.17 (m, 2H), 4.04(m, 4H), 2.75 (m, 1H), 2.38 (m, 2H), 2.09 (m, 1H), 1.91 (m, 1H), 1.65(m, 8H) 1.34 (d, J=6.3 Hz, 6H), 1.04 (s, 9H). ³¹P (121.4 MHz, CD₃OD):

31.453. LC/MS=837 (M⁺+1)

Example 15 Preparation of Compound 15

The[(1-Benzyloxycarbonylamino-2-vinyl-cyclopropyl)-ethoxy-phosphinoyl]-aceticacid from example 11 (340 mg, 0.92 mmol) was suspended in 5 mL of DMF.HATU (1.04 g, 2.76 mmol), ammonium chloride (123 mg, 2.32 mmol),followed by NMM (910 μl, 8.28 mmol) was added. After 2 hours, thereaction was concentrated and partitioned with EtOAc and H₂O. Theaqueous layer was extracted 3× with EtOAc. The organic layer was driedover MgSO4, filtered and concentrated. The product,(1-benzyloxycarbonylamino-2-vinyl-cyclopropyl)-carbamoylmethyl-phosphinicacid ethyl ester as brown oil (214 mg, 64%) was used as crude.

The crude(1-benzyloxycarbonylamino-2-vinyl-cyclopropyl)-carbamoylmethyl-phosphinicacid ethyl ester (107 mg, 0.29 mmol) was suspended in 1 mL of CH₃CN andcooled to 0° C. Iodotrimethylsilyl (TMSI) (208 μl, 1.46 mmol) was addedand the solution was warmed to rt. After 45 minutes, the solution wascooled again to 0° C. and triethylamine (1 mL, 7.33 mmol) and 2 mL ofMeOH. The solution was warmed to rt and stirred for an additional 20minutes. The solution was concentrated, azeotroped 2× with toluene andput on high vacuum for 30 minutes. The residue was coupled with VII (94mg, 0.14 mmol), HATU (133 mg, 0.35 mmol) and NMM (77 μl, 0.70 mmol). Themixture was purified via Gilson HPLC to obtain 15 (15.4 mg, 13%) as ayellow solid. ^(1H) NMR (300 MHz, CD₃OD): d 8.23 (d, J=9.5 Hz, 1H), 8.20(s, 1H), 7.79 (s, 2H), 7.33 (d, J=8.8 Hz, 1H), 5.95 (m, 1H), 5.78 (s,1H), 5.22 (d, J=9.6 Hz, 2H), 5.13 (d, J=9.0 Hz, 2H), 4.63 (m, 2H), 4.45(bs, 1H), 4.20 (s, 2H), 4.05 (s, 3H), 3.25 (m, 1H), 2.80 (m, 2H), 2.45(m, 1H), 2.15 (m, 1H), 1.62 (m, 6H), 1.38 (d, 6H), 1.05 (s, 9H). ³¹P(121.4 MHz, CD₃OD):

37.283

Example 16 Preparation of Compound 16

Examples 16-23 were prepared using Grignard reagents. A detailedprocedure is described in example 19. ¹H NMR (300 MHz, CD₃CN): d 8.25(m, 1H), 8.20 (m, 1H), 7.63 (s, 1H), 7.41 (m, 1H), 7.23 (m, 1H), 6.81(br, 1H), 6.37 (m, 1H), 6.02 (m, 3H), 5.60 (br, 1H), 5.13 (m, 1H), 4.98(m, 1H), 4.60 (m, 2H), 4.19 (m, 2H), 4.05 (m, 2H), 4.00 (s, 3H), 2.70(m, 1H), 2.43 (m, 1H), 1.65 (m, 8H) 1.38 (d, 6H), 1.21 (m, 1H), 1.05 (s,9H). ³¹P (121.4 MHz, CD₃CN):

30.642

LC/MS=809 (M⁺+1)

Example 17 Preparation of Compound 17

¹H NMR (300 MHz, CD₃OD): d 8.27 (d, J=9.6 Hz, 1H), 8.18 (s, 1H), 7.75(s, 2H), 7.³¹ (m, 1H), 6.67 (m, 1H), 5.93 (m, 2H), 5.77 (bs, 1H), 5.24(d, J=17.1 Hz, 1H), 5.07 (d, J=11.4 Hz, 1H), 4.63 (m, 2H), 4.43 (bs,1H), 4.20 (m, 2H), 4.04 (m, 5H), 3.23 (m, 1H), 2.76 (m, 1H), 2.40 (m,1H), 2.08 (m, 1H), 1.90 (m, 3H), 1.72-1.40 (m, 8H), 1.38 (d, 6H), 1.05(m, 9H). ³¹P (121.4 MHz, CD₃OD):

33.223. LC/MS=823 (M⁺+1)

Example 18 Preparation of Compound 18

¹H NMR (300 MHz, CD₃OD): d 8.27 (d, J=9.6 Hz, 1H), 8.17 (s, 1H), 7.74(s, 2H), 7.32 (m, 1H), 6.63-6.41 (m, 1H), 5.98 (m, 2H), 5.77 (bs, 1H),5.24 (d, J=17.1 Hz, 1H), 5.07 (d, J=11.4 Hz, 1H), 4.63 (m, 2H), 4.43(bs, 1H), 4.17 (m, 2H), 4.07 (m, 4H), 2.75 (m, 1H), 2.42 (m, 1H), 2.10(m, 1H), 2.07 (m, 3H), 1.72-1.40 (m, 8H), 1.34 (d, J=6.3 Hz, 6H), 1.04(m, 9H). ³¹P (121.4 MHz, CD₃OD):

33.781. LC/MS=823 (M⁺+1)

Example 19 Preparation of Compound 19

Intermediate III (1.0 g, 3.1 mmol) was dissolved in toluene (20 mL).This solution was cooled to 0° C. and (COCl)₂ (1.6 g, 12.4 mmol) wasadded in a drop-wise fashion. DMF (45 mg, 0.62 mmol) was then added. Thereaction was run for 2 h at 0° C. and determined to be complete by ³¹PNMR. The reaction was concentrated to an orange-yellow oil and thenplaced under high vacuum for 1 h. The resulting residue was dissolved inTHF (6.4 mL) and this solution was cooled to −78° C. A 1.0M solution ofcis-1-butenemagnesium bromide in THF (9.1 mL, 9.1 mmol) was addeddrop-wise. The reaction mixture was warmed to rt and stirred for 3hours. The reaction was quenched at 0° C. by the addition of sat.NH₄Cl_((aq.)). The organic phase was diluted with EtOAc and extractedwith sat. NH₄Cl_((aq.)) and brine. The organic phase was dried overMgSO₄. Concentration of the filtrate after vacuum filtration removal ofthe MgSO₄ yielded an orange oil from which(1-Benzyloxycarbonylamino-2-vinyl-cyclopropyl)-but-1-enyl-phosphinicacid ethyl ester was isolated by column chromatography (SiO₂, 100%EtOAc) as a clear oil (100 mg, 8% over 2 steps). ¹H NMR (300 MHz,CDCl₃): d 7.33 (m, 5H), 6.60-6.35 (m, 1H) 6.18-5.83 (m, 1H), 5.68 (m,1H), 5.38 (m, 2H), 5.10 (m, 3H), 4.05 (m, 2H), 2.57 (m, 2H), 2.01 (m,1H), 1.78 (m, 1H), 1.50 (m, 1H), 1.23 (m, 3H), 1.00 (m, 3H).

³¹P (121.4 MHz, CDCl₃):

37.397, 35.875 diastereomers

The phosphinate (100 mg, 0.275 mmol) was suspended in 1 mL of CH₃CN andcooled to 0° C. Iodotrimethylsilane (TMSI) (190 μL, 1.38 mmol) was addedand the solution was warmed to rt. After 45 minutes, the solution wascooled again to 0° C. and triethylamine (0.5 mL, 3.6 mmol) and 0.5 mL ofMeOH was added to the reaction. The reaction was warmed to rt andstirred for an additional 20 minutes. The reaction was concentrated,azeotroped twice with toluene and put on high vacuum for 30 minutes. Thesolid was coupled to VII to give compound 19 after reverse phase HPLCpurification. ¹H NMR (300 MHz, CD₃OD): d 8.25 (d, J=9.0 Hz, 1H), 8.19(s, 1H), 7.73 (s, 2H), 7.35 (m, 1H), 6.52-6.28 (m, 1H), 5.95 (m, 2H),5.77 (s, 1H), 5.24 (d, J=17.9 Hz, 1H), 5.06 (d, J=11.1 Hz, 1H), 4.65 (m,4H), 4.44 (bs, 1H), 4.20 (m, 2H), 4.04 (m, 4H), 2.76 (m, 1H), 2.52 (m,3H), 2.43 (m, 1H), 2.13 (m, 1H), 1.62-1.35 (m, 10H) 1.38 (d, J=6.3 Hz,6H), 1.03 (m, 12H).

³¹P (121.4 MHz, CD₃OD):

34.248. LC/MS=837 (M⁺+1)

Example 20 Preparation of Compound 20

¹H NMR (300 MHz, CD₃OD): d 8.27 (d, J=9.6 Hz, 1H), 8.18 (s, 1H), 7.74(s, 2H), 7.32 (m, 1H), 5.97 (m, 1H), 5.72 (m, 2H), 5.24 (d, J=16.5 Hz,1H), 5.07 (d, J=10.5 Hz, 1H), 4.66 (m, 2H), 4.43 (bs, 1H), 4.20 (m, 2H),4.06 (m, 5H), 2.75 (m, 1H), 2.43 (m, 1H), 2.11 (m, 4H), 1.91 (m, 3H),1.72-1.40 (m, 10H), 1.38 (d, J=6.2 Hz, 6H), 1.04 (m, 9H). ³¹P (121.4MHz, CD₃OD):

33.786

LC/MS=837 (M⁺+1).

Example 21 Preparation of Compound 21

¹H NMR (300 MHz, CD₃OD): d 8.25 (d, J=9.0 Hz, 1H), 8.15 (s, 1H), 7.74(m, 1H), 7.68 (m, 1H), 7.57 (m, 2H), 7.40 (m, 4H), 6.57 (m, 1H), 5.98(m, 1H), 5.68 (bs, 1H), 5.27 (d, J=17.1 Hz, 1H), 5.10 (d, J=9.0 Hz, 1H),4.63 (m, 2H), 4.44 (bs, 1H), 4.18 (m, 2H), 4.04 (m, 4H), 3.31 (m, 1H),2.70 (m, 1H), 2.38 (m, 1H), 2.15 (m, 1H), 1.72-1.40 (m, 8H), 1.38 (d,J=6.3 Hz, 6H), 1.03 (m, 9H). ³¹P (121.4 MHz, CD₃OD):

33.372. LC/MS=885 (M⁺+1)

Example 22 Preparation of Compound 22

¹H NMR (300 MHz, CD₃OD) d 8.25 (d, J=9 Hz, 1H), 8.17 (s, 1H), 7.74 (d,J=2.1 Hz, 1H), 7.68 (s, 1H), 7.55 (d, J=6.9 Hz, 2H), 7.2-7.5 (m, 4H),6.05 (dt, J=9.6, 17.1 Hz), 5.71 (s, 1H), 5.27 (d, J=17.4 Hz, 1H), 5.09(d, J=9.6 Hz, 1H), 4.7 (t, J=8.7 Hz, 1H), 4.6 (d, J=12.6 Hz, 1H), 4.51(s, 1H), 4.06-4.2 (brm, 3H), 4.04 (s, 3H), 2.74 (dd, J=7.8, 13.8 Hz,1H), 2.57 (m, 1H), 2.28 (m, 1H), 1.36-1.9 (brm, 10H), 1.33 (d, J=6.3 Hz,6H), 1.04 (s, 9H). ³¹P NMR (121.4 MHz, CD₃OD) d 10.2. LC/MS=883 (M⁺+1)

Example 23 Preparation of Compound 23

The phosphonous acid IV (363 mg, 1.17 mmol) was suspended in 5 mL of THFand cooled to −40° C. 1N NaN(TMS)₂ (1.41 mL, 1.41 mmol) was addeddropwise over 15 minutes followed by 1-bromo-3-methylbut-2-ene (164 μl,1.41 mmol) in 1 mL of THF. The solution was stirred from −40° C. to rtover 45 minutes. The reaction was diluted with EtOAc and quenched with20 mL of 1N HCl. The organic layer was washed with brine, dried overMgSO₄, filtered and concentrated. The crude material was purified usinga CombiFlash Chromatography System employing a gradient of 30%EtOAc/Hexane to 100% EtOAc to obtain(1-benzyloxycarbonylamino-2-vinyl-cyclopropyl)-(3-methyl-but-2-enyl)-phosphinicacid ethyl ester (219 mg, 50%) as a brown oil. This oil (135 mg, 0.36mmol) was suspended in 1 mL of CH₃CN and cooled to 0° C.Iodotrimethylsilane (254 μl, 1.79 mmol) was added and the solution waswarmed to rt. After 45 minutes, the solution was cooled again to 0° C.and triethylamine (1 mL, 7.33 mmol) and 2 mL of MeOH was added to thereaction. The reaction was warmed to rt and stirred for an additional 20minutes. The reaction was concentrated, azeotroped 2× with toluene andput on high vacuum for 30 minutes. The crude was coupled withintermediate VII to give compound 23 after HPLC purification. ¹H NMR(300 MHz, CD₃OD): d 8.30 (d, J=9.5 Hz, 1H), 8.20 (s, 1H), 7.79 (s, 2H),7.30 (m, 4H), 5.95 (m, 1H), 5.80 (s, 1H), 5.25 (d, J=9.6 Hz, 2H), 5.13(d, J=9.0 Hz, 2H), 4.75 (m, 2H), 4.45 (bs, 1H), 4.20 (s, 2H), 4.05 (s,3H), 3.33 (s, 2H), 2.80 (m, 1H), 2.52 (m, 1H), 2.15 (m, 1H), 1.62 (m,6H), 1.38 (d, 6H), 1.05 (s, 9H). ³¹P (121.4 MHz, CD₃OD):

42.837

Example 24 Preparation of Compound 24

A suspension of sodium borohydride (82 mg, 2.17 mmol) and[(1-Benzyloxycarbonylamino-2-vinyl-cyclopropyl)-ethoxy-phosphinoyl]-aceticacid ethyl ester from example 91 (344 mg, 0.87 mmol) in THF (3.5 mL) wasstirred at 50° C. as MeOH (710 μL) was added dropwise over 20 minutes.After 20 minutes at 50° C., the reaction was concentrated and theresulting residue in ethyl acetate (15 mL) was washed with H₂O andbrine. The aqueous layer was extracted with ethyl acetate and thecombined organic layers were dried (MgSO₄) and concentrated to yieldalcohol (282 mg, 91.8%). The product was used without furtherpurification.

¹H NMR (300 MHz, CDCl₃): d 7.35 (s, 5H), 5.99 (m, 1H), 5.64 (d, 1H),5.38 (dd, 1H), 5.07 (s, 2H), 4.12 (m, 2H), 3.91 (m, 2H), 2.96 (bs, 1H),2.18 (m, 3H), 1.76 (m, 1H), 1.62 (m, 1H), 1.50 (m, 1H), 1.26 (m, 3H).³¹P (121.4 MHz, CDCl₃):

52.755, 49.793.

The alcohol (112 mg, 0.32 mmol) was suspended in 1 mL of CH₃CN andcooled to 0° C. Iodotrimethylsilane (225 μl, 1.58 mmol) was added andthe solution was warmed to rt. After 45 minutes, the solution was cooledagain to 0° C. and triethylamine (1 mL, 7.33 mmol) and 2 mL of MeOH wasadded to the reaction. The reaction was warmed to rt and stirred for anadditional 20 minutes. The reaction was concentrated, azeotroped 2× withtoluene and put on high vacuum for 30 minutes. The solid (104 mg, 0.16mmol) was suspended in 1 mL of DMF. HATU (152 mg, 0.40 mmol), VII (61mg, 0.32 mmol), and NMM (88 μl, 0.80 mmol) was added. The solution wasstirred overnight at rt. The mixture was purified by reverse phase HPLCto obtain 24 (33.3 mg, 25%) as a yellow solid.

¹H NMR (300 MHz, CD₃OD): d 8.23 (d, J=9.5 Hz, 1H), 8.20 (s, 1H), 7.79(s, 2H), 7.33 (d, J=8.8 Hz, 1H), 5.95 (m, 1H), 5.78 (s, 1H), 5.22 (d,J=9.6 Hz, 2H), 5.13 (d, J=9.0 Hz, 2H), 4.63 (m, 2H), 4.45 (bs, 1H), 4.20(m, 3H), 4.05 (s, 3H), 3.83 (m, 2H), 2.80 (m, 1H), 2.78 (s, 3H), 2.45(m, 1H), 2.20 (m, 1H), 2.15 (m, 1H), 1.62 (m, 2H), 1.50 (min, 6H) 1.38(d, 6H), 1.05 (s, 9H).

³¹P (121.4 MHz, CD₃OD):

45.011

Example 25 Preparation of Compound 25

¹H NMR (300 MHz, CD₃OD) d 8.29 (d, J=9.6 Hz, 1H), 8.18 (s, 1H), 7.75 (m,2H), 7.32 (dd, J=3, 9.3 Hz), 6.00 (dt, J=10.2, 16.5 Hz, 1H), 5.78 (s,1H), 5.27 (d, J=15.6 Hz), 5.10 (d, J=12 Hz, 1H), 4.64 (m, 2H), 4.44(1H), 4.17 (m, 2H), 4.08 (m, 1H), 4.05 (s, 3H), 2.76 (dd, J=6.6, 13.5Hz, 1H), 2.45 (m, 1H), 2.32 (m, 1H), 2.09 (m, 2H), 1.37-1.65 (brm, 16H),1.34 (d, J=6.3 Hz, 6H), 1.05 (s, 9H). ³¹P NMR (121.4 MHz, CD₃OD) d 45.7.LC/MS=854.7 (M⁺+1)

Example 26 Preparation of Compound 26

Intermediate IV (467 mg, 1.5 mmol) was dissolved in 5.0 mL of dry DCM.DIEA (523 μl, 3.0 mmol) and 381 μl (3.0 mmol) of TMSCl were addedsequentially and the reaction then stirred at rt for 5 min. DIEA (523μl, 3.0 mmol) and 209 μl (3.0 mmol) of bromoacetonitrile were thenadded. The reaction was warmed to 40° C. and stirred overnight. Thereaction was then diluted with ethyl acetate and concentrated to removeDCM. The organic phase was then washed with sat. NH₄Cl, water, andbrine. The organic phase was dried over MgSO₄. Concentration of thefiltrate after vacuum filtration removal of the MgSO₄ yielded an orangeoil from which the product was isolated by column chromatography (SiO₂,neat ethyl acetate) as a clear oil (190 mg, 37%). ¹H NMR (300 MHz,CDCl₃) d 7.35 (s, 5H), 5.81 (m, 1H), 5.60-5.26 (m, 2H), 5.11 (s, 2H),4.23 (m, 2H), 2.99 (m, 2H), 2.18 (m, 1H), 1.85-1.70 (m, 1H), 1.65-1.47(m, 1H), 1.35 (m, 3H). ³¹P NMR (121.4 MHz, CDCl₃) d 39.04, 36.33.LC/MS=370 (M⁺+1)

Deprotection and coupling to dipeptide VII as described above gave 26(60 mg 40%). ¹H NMR (300 MHz, CD₃OD) 8.30 (d, J=9.3 Hz, 1H), 8.23 (s,1H), 7.74 (s, 2H), 7.30 (d, J=2.1, 8.7 Hz, 1H), 5.90 (m, 2H), 5.76 (b,1H), 5.20 (d, J=17.4 Hz, 1H), 5.05 (d, J=11.1 Hz, 1H), 4.61 (m, 2H),4.55 (s, 1H), 4.18 (m, 2H), 4.11 (s, 1H), 4.04 (s, 3H), 3.0 (m, 2H),2.70 (m, 1H), 2.60 (m, 1H), 2.00 (m, 1H), 1.41-1.78 (m, 8H), 1.34 (d,J=6.3 Hz, 6H), 1.04 (s, 9H)

³¹P NMR (121.4 MHz, CD₃OD) d 24.48

LC/MS=822 (M⁺+1)

Example 27 Preparation of Compound 27

A solution of the phosphonous acid IV (436 mg, 1.40 mmol), Hunig's base(593 μL, 3.40 mmol), and chlorotrimethylsilane (378 μL, 3.12 mmol) inCH₂Cl₂ (5 mL) was stirred at r.t. for 1 h. After chloro(methoxy)methane(220 μL, 3.17 mmol) was added, the solution was heated at 40° C. for 2h. The solution was concentrated and the residue in ethyl acetate (30mL) was washed with H₂O (30 mL×2). The aqueous fractions were extractedwith ethyl acetate (30 mL), and the combined organic fractions weredried (MgSO₄) and concentrated. The residue was purified by columnchromatography using hexane:ethyl acetate as eluent to obtain 27 (297mg, 60%). ¹H NMR (300 MHz, CDCl₃): d 7.35 (s, 5H), 6.00 (m, 1H), 5.44(m, 2H), 5.15 (m, 1H), 5.07 (s, 2H), 4.18 (m, 2H), 3.87 (m, 1H), 3.77(d, J=6.6 Hz, 2H), 3.43 (s, 3H), 2.20 (m, 1H), 2.07 (m, 1H), 1.80 (m,1H), 1.64 (m, 1H), 1.48 (m, 1H), 1.28 (m, 3H). ³¹P (121.4 MHz, CDCl₃):

44.0099, 43.403, 40.648, 40.353.

Deprotection and coupling to dipeptide VII as described above gave 27.

LC/MS=827 (M⁺+1)

Example 28 Preparation of Compound 28

Compound IV (1.64 g, 5.³¹ mmol) was dissolved in CH₂Cl₂ (60 mL) andcooled to 0° C. Diisopropylethylamine (1.96 mL) was added and stirredfor 15 minutes. Chlorotrimethylsilane (1.40 mL) was added dropwise. Thereaction mixture was warmed to r.t. and stirred for 1 h. Ethylbromoacetate (2.92 mL) was added and the reaction was heated to 45° C.overnight. The reaction mixture was cooled to rt, diluted with CH₂Cl₂,washed with aqueous NH₄Cl, dried with Na₂SO₄, and concentrated. Thecrude product was purified by column chromatography to give 1.15 g of[(1-Benzyloxycarbonylamino-2-vinyl-cyclopropyl)-ethoxy-phosphinoyl]-aceticacid ethyl ester in 55% yield.

To a solution of ester (679 mg, 1.72 mmol) in toluene (25 mL) at −78° C.was added 1.0 M DIBAL in CH₂Cl₂ (6.6 mL, 6.6 mmol) and stirred for 2 h.The mixture was poured into ice cold 6 N HCl (100 mL), extracted withEtOAc, and concentrated. The residue was re-dissolved in CH₂Cl₂,insoluble material was removed by a filtration through celite, and thefiltrate was concentrated to give a colorless oil. The oil was dissolvedin CH₂Cl₂ (20 mL) and then AcOH (0.52 mL), trifluoroethylamine (260 mg),and sodium triacetoxyborohydride (730 mg) were added sequentially. Themixture was stirred at r.t. for 16 h. The reaction was partitionedbetween CH₂Cl₂ and saturated NaHCO₃. The organic layer was concentratedand purified by column chromatography to give 310 mg of phosphinate asan oil.

To a solution of phosphinate in CH₃CN (1 mL) at 0° C. was addediodotrimethylsilane (0.03 mL). The reaction mixture was warmed to rt,stirred for 0.5 h, and cooled to 0° C. Triethylamine (0.2 mL) was addedfollowed by MeOH (2 mL) and the reaction was warmed to rt. The mixturewas concentrated and dried under vacuum to give 23 mg of amine as crudeproduct.

Acid VII (35 mg) was dissolved in DMF (0.8 mL). HATU (30 mg) was addedand the mixture was cooled to 0° C. DIPEA (0.04 mL) was added and themixture was stirred at r.t. for 1 h. A solution of the amine in CH₂Cl₂(2 mL) was added and stirred for 1 h. The reaction was quenched with H₂Oand CH₂Cl₂ was removed in vacuo. The non-volatile residue was purifiedby HPLC to give 19.9 mg of compound 28. ¹H NMR (300 MHz, CD₃OD): d 8.28(d, J=9.3 Hz, 1H), 8.18 (s, 1H), 7.74 (s, 2H), 7.30 (dd, J=2.4, 9.0 Hz,1H), 5.97 (m, 1H), 5.79 (brs, 1H), 5.23 (d, J=17.7 Hz, 1H), 5.06 (d,J=11.7 Hz, 1H), 4.65 (m, 2H), 4.46 (brs, 1H), 4.15 (m, 2H), 3.90-4.10(m, 6H), 3.55 (m, 1H), 3.39 (m, 1H), 2.80 (m, 1H), 2.45 (m, 1H), 2.12(m, 3H), 1.4-1.7 (m, 10H), 1.34 (d, J=6.3 Hz, 6H), 0.95-1.15 (brs, 9H).

Example 29 Preparation of Compound 29

A solution of compound IV (149 mg, 0.482 mmol) in THF (2.41 mL) wascooled to −40° C. To the solution was added a solution of 1M NaHMDS inTHF (0.578 mL) and the resulting mixture was stirred for 30 minutes andthen 2-bromoethylbenzene (107 mg, 0.578 mmol) was added. The resultingsolution was stirred for 2 additional hours until all of the startingmaterials was consumed as determined by LCMS. The reaction was worked upby removal of the solvent in vacuo. The residue was dissolved in EtOAcand washed with saturated aqueous NH₄Cl. The organic layer was dried andthe product was purified using silica gel chromatography to give 74 mgof the product as a clear oil.

EI MS (m/z) 436.1 [M+Na].

To a solution of benzyl(1S,2S)-1-((S)-ethoxy-(phenethyl)phosphoryl)-2-vinylcyclopropylcarbamate(72 mg, 0.174 mmol) in dry acetonitrile (1.74 mL) was added TMSI (0.124mL, 0.87 mmol). The reaction was stirred at ambient temperature for 1hour until LCMS analysis indicated completion of the reaction. Themixture was quenched by addition of TEA (0.243 mL, 1.74 mmol) followedby MeOH (10 mL). The residue was dried and used without furtherpurification.

EI MS (m/z) 252.3 [MH⁺], 274.1 [M+Na].

A solution of(S)-((1S,2S)-1-amino-2-vinylcyclopropyl)(phenethyl)phosphinic acid (43mg, 0.171 mmol), carboxylic acid VII (112 mg, 0.171 mmol) in a 1:1solution of DMF and CH₂Cl₂ (1.7 mL) was stirred with HATU (98 mg, 0.256mmol) and DIEA (0.119 mL, 0.685 mmol) for 1 hour until the reaction wascomplete. The product was purified by reverse phase HPLC (ACN, 0.05%TFA-H₂O, 0.05% TFA) to provide the desired product. ¹H NMR (300 MHz,CD₃OD) d 8.27 (d, 1H, J=9 Hz), 8.16 (s, 1H), 7.75-7.71 (m, 2H), 7.30 (d,1H, J=11 Hz), 7.27-7.22 (m, 5H), 6.01 (dt, 1H, J=17, 10 Hz), 5.75 (br s,1H), 5.28 (d, 1H, J=17 Hz), 5.11 (d, 1H, J=11 Hz), 4.68-4.58 (m, 2H),4.44 (br s, 1H), 4.22-4.10 (m, 2H), 4.04 (s, 3H), 3.05-2.83 (m, 2H),2.82-2.70 (m, 1H), 2.48-2.37 (m, 1H), 2.18-2.03 (m, 3H), 1.68-1.40 (m,10H), 1.33 (d, 6H, J=6 Hz), 0.99 (s, 9H); ³¹P (121.4 MHz, CD₃OD) d 47.2;EI MS (m/z) 887.4 [MH⁺].

Example 30 Preparation of Compound 30

Examples 30 through 33 were prepared by the same method as example 29.

Preparatory reverse phase HPLC afforded compound 30 (10 mg, 33%), ayellow solid. ¹H NMR (300 MHz, CD₃OD): d 8.27 (d, J=9.4 Hz, 1H), 8.16(s, 1H), 7.74 (s, 1H), 7.72 (s, 1H), 7.32 (m, 3H), 7.23 (m, 2H) 6.00 (m,1H), 5.75 (s, 1H), 5.27 (m, 1H), 5.10 (m, 1H) 4.64 (m, 2H), 4.46 (m,1H), 4.16 (m, 3H), 4.04 (s, 3H), 3.10 (m, 2H), 2.76 (m, 1H), 2.43 (m,1H), 2.10 (m, 3H), 1.60 (m, 8H) 1.34 (m, 6H), 1.02 (s, 9H). ³¹P (121.4MHz, CD₃OD):

44.597

LC (6 minute run, r.t.=3.82 min) MS (921.6, M+1)

Example 31 Preparation of Compound 31

Preparatory reverse phase HPLC purification afforded compound 31 (23 mg,47%) as a yellow solid. ¹H NMR (300 MHz, CD₃OD): d 8.22 (d, J=9.2 Hz,1H), 8.11 (s, 1H), 7.70 (s, 1H), 7.68 (s, 1H), 7.25 (m, 3H), 6.99 (m,2H) 5.98 (min, 1H), 5.70 (s, 1H), 5.21 (min, 1H), 5.05 (m, 1H) 4.58 (m,2H), 4.40 (s, 1H), 4.11 (m, 2H), 3.99 (s, 3H), 2.91 (m, 2H), 2.70 (m,1H), 2.38 (m, 1H), 2.08 (m, 3H), 1.50 (m, 8H) 1.29 (m, 6H), 0.93 (s,9H). ³¹P (121.4 MHz, CD₃OD):

44.896. LC (6 minute run, r.t.=3.70 min) MS (905.5, M+1)

Example 32 Preparation of Compound 32

Preparatory reverse phase HPLC purification afforded compound 32 (85 mg,65%) as a yellow solid. ¹H NMR (300 MHz, CD₃OD): d 8.215 (d, J=9.3 Hz,1H), 8.11 (s, 1H), 7.70 (s, 1H), 7.69 (s, 1H), 7.25 (m, 2H), 7.025 (m,4H) 5.95 (m, 1H), 5.69 (s, 1H), 5.22 (m, 1H), 5.06 (m, 1H) 4.59 (m, 2H),4.40 (s, 1H), 4.10 (m, 2H), 3.99 (s, 3H), 2.90 (m, 2H), 2.70 (m, 1H),2.36 (m, 1H), 2.26 (s, 3H) 2.10 (m, 3H), 1.50 (m, 8H) 1.29 (m, 6H), 0.93(s, 9H). ³¹P (121.4 MHz, CD₃OD):

45.420. LC (6 minute run, r.t.=3.77 min) MS (902.6, M+1)

Example 33 Preparation of Compound 33

Preparatory reverse phase HPLC purification afforded compound 33 (70 mg,55%) as a yellow solid. ¹H NMR (300 MHz, CD₃OD): d 8.25 (d, J=9.1 Hz,1H), 8.17 (s, 1H), 7.74 (m, 2H), 7.3 (m, 1H), 7.21 (m, 2H), 7.04 (m, 1H)5.95 (m, 1H), 5.75 (bs, 1H), 5.25 (m, 1H), 5.10 (m, 1H) 4.60 (m, 2H),4.40 (bs, 1H), 4.13 (m, 2H), 4.04 (s, 3H), 3.09 (m, 2H), 2.70 (m, 1H),2.42 (m, 1H), 2.10 (m, 3H), 1.50 (m, 8H) 1.33 (m, 6H), 0.97 (s, 9H). ³¹P(121.4 MHz, CD₃OD):

44.588. LC (6 minute run, r.t.=4.22 min) MS (940.3, M+1).

Example 34 Preparation of Compound 34

Intermediate IV (1.08 g, 3.5 mmol) was dissolved in CH₂Cl₂ (40 mL) andcooled to 0° C. Diisopropylethylamine (950 mg, 7.35 mmol) was added andstirred for 15 minutes. Chlorotrimethylsilane (800 mg, 7.35 mmol) wasadded dropwise. The reaction mixture was warmed to r.t. and stirred for1 h. Chloromethylsulfanyl-benzene (2.77 g, 17 mmol) was added and thereaction was heated to 45° C. overnight. The reaction mixture was cooledto rt, diluted with CH₂Cl₂, washed with aqueous NH₄Cl, dried withNa₂SO₄, and concentrated. The crude product was purified by combi-flashto give 222 mg of phosphinate.

To a solution of phosphinate obtained above (222 mg, 0.52 mmol) in CH₃CN(1 mL) at 0° C. was added iodotrimethylsilane (0.36 mL, 2.58 mmol). Thereaction mixture was warmed to r.t. and stirred for 30 minutes and thencooled to 0° C. 2,6-Lutidine (0.3 mL) and MeOH (0.6 mL) were added andstirred for 10 minutes. The solvent was concentrated and the residue wasco-evaporated with toluene (5 mL), and dried under vacuum for 20 minutesto give crude amine. Coupling with acid VII (168 mg, 0.26 mmol) provided150 mg of compound 34. ¹H NMR (300 MHz, CD₃OD):

8.27 (d, J=9.6 Hz, 1H), 8.20 (s, 1H), 7.72 (s, 2H), 7.41 (d, J=7.8 Hz,1H), 7.27 (m, 3H), 7.15 (d, J=7.2 Hz, 1H), 5.94 (m, 1H), 5.74 (s, 1H),5.28 (d, J=17.1 Hz, 1H), 5.11 (d, J=11.1 Hz, 1H), 4.63 (m, 2H), 4.48 (s,1H), 4.16 (m, 2H), 3.36 (m, 2H), 2.74 (m, 1H), 2.46 (m, 1H), 2.10 (m,1H), 1.70-1.40 (m, 8H), 1.30 (m, 6H), 0.97 (s, 9H) ³¹P (121.4 MHz,CDCl₃): d 39.174

LC/MS=905.20 (M⁺+1)

Example 35 Preparation of Compound 35

¹H NMR (300 MHz, CD₃OD) d 8.29 (d, J=9.3 Hz, 1H), 8.18 (s, 1H), 7.75 (s,2H), 7.17-7.34 (brm, 6H), 5.96 (dt, J=9.9, 17.1 Hz, 1H), 5.79 (s, 1H),5.27 (d, J=17.1 Hz, 1H), 5.09 (d, J=10.2 Hz, 1H), 4.69 (m, 2H), 4.46 (s,1H), 4.07-4.2 (brm, 3H), 4.05 (s, 3H), 3.29 (d, J=15.6 Hz, 2H), 2.78(dd, J=7.5, 14.1 Hz, 1H), 2.48 (m, 1H), 2.11 (m, 1H), 1.38-1.7 (brm,10H), 1.34 (d, J=6.3 Hz, 6H), 1.02 (s, 9H). ³¹P NMR (121.4 MHz, CD₃OD) d43.3. LC/MS=872.7 (M⁺+1), 894.5 (M⁺+Na)

Example 36 Preparation of Compound 36

The phosphonous acid IV (409 mg, 1.32 mmol) was suspended in 2.5 mL ofCDCl₃. The air was removed from the reaction flask by vacuum andreplaced with N₂. Hunig's Base (552 μl, 3.16 mmol) followed bychlorotrimethylsilyl (368 μl, 2.90 mmol) was added. After 5 minutes,1-(bromomethyl)-2-fluorobenzene (334 μl, 2.77) was added and thesolution was heated at 40° C. After 4 hours, the reaction wasconcentrated. The residue was partitioned with EtOAc and H₂O and washedwith H₂O. The organic layer was dried over MgSO₄, filtered andconcentrated. The crude material was purified using a CombiFlashChromatography System using a gradient of 50% EtOAc/Hex to 100% EtOAc toobtain(1-benzyloxycarbonylamino-2-vinyl-cyclopropyl)-(2-fluoro-benzyl)-phosphinicacid ethyl ester (142.8 mg, 26%) as a brown oil. ^(1H) NMR (300 MHz,CDCl₃): d 7.29-7.48 (m, 6H), 7.16-7.29 (m, 1H), 7.16-6.98 (m, 2H), 6.06(dt, 0.4H, J=17.1 and 10.2 Hz), 5.76 (dt, 0.6H, J=17.1 and 9.9 Hz),5.28-5.41 (m, 0.6H), 4.96-5.22 (m, 3.8H), 4.90 (d, 0.6H, J=9.9 Hz),3.9-4.17 (m, 2H), 3.05-3.53 (m, 2H), 2.11-2.26 (m, 0.4H), 1.91-2.05 (m,0.6H), 1.70-1.82 (m, 1.4H), 1.50-1.60 (m, 0.6H), 1.05-1.32 (m, 4H). ³¹P(121.4 MHz, CDCl₃): d 46.333, (0.4P), 49.339 (0.6P). ¹⁹F (121.4 MHz,CDCl₃): d −112.9³¹ (0.6F), −118.³¹5, 0.4F).

The residue (142.8 mg, 0.34 mmol) was suspended in 1 mL of CH₃CN andcooled to 0° C. Iodotrimethylsilyl(TMSI) (243 μl, 1.71 mmol) was addedand the solution was warmed to rt. After 45 minutes, the solution wascooled again to 0° C. and triethylamine (1 mL, 7.33 mmol) and 2 mL ofMeOH. The solution was warmed to rt and stirred for an additional 20minutes. The solution was concentrated, azeotroped 2× with toluene andput on high vacuum for 30 minutes. The crude was coupled with acid VII(148 mg, 0.23 mmol), HATU (218 mg, 0.58 mmol) and NMM (1261, 1.15 mmol)to give 36 (122 mg, 60%) as a yellow solid after Gilson HPLCpurification. ¹H NMR (300 MHz, CD₃OD): d 8.30 (d, J=9.5 Hz, 1H), 8.20(s, 1H), 7.79 (s, 2H), 7.42 (t, 1H), 7.35 (d, J=8.8 Hz, 1H), 7.22 (m,1H), 7.15 (m, 2H), 5.95 (m, 1H), 5.78 (s, 1H), 5.35 (d, J=9.6 Hz, 2H),5.15 (d, J=9.0 Hz, 2H), 4.75 (m, 2H), 4.45 (bs, 1H), 4.20 (s, 2H), 4.05(s, 3H), 3.33 (m, 2H), 2.80 (m, 1H), 2.52 (m, 1H), 2.20 (m, 1H), 1.62(m, 6H), 1.38 (d, 6H), 1.05 (s, 9H). ³¹P (121.4 MHz, CD₃OD):

42.259.

Example 37 Preparation of Compound 37

The phosphonous acid IV (350 mg, 1.13 mmol) was suspended in 2.5 mL ofCDCl₃. The air was removed from the reaction flask by vacuum andreplaced with N₂. Hunig's Base (472 μl, 2.71 mmol) followed byChlorotrimethylsilyl (³¹5 μl, 2.48 mmol) was added. After 5 minutes,1-(bromomethyl)-3-fluorobenzene (449 mg, 2.37) in 500 μl of CDCl₃ wasadded and the solution was heated at 40° C. After 4 hours, the reactionwas concentrated. The residue was partitioned with EtOAc and H₂O andwashed with H₂O. The organic layer was dried over MgSO₄, filtered andconcentrated. The crude material was purified using a CombiFlashChromatography System using a gradient of 50% EtOAc/Hex to 100% EtOAc toobtain(1-benzyloxycarbonylamino-2-vinyl-cyclopropyl)-(3-fluoro-benzyl)-phosphinicacid ethyl ester (110 mg, 23%) as a brown oil.

The residue (110 mg, 0.26 mmol) was suspended in 1 mL of CH₃CN andcooled to 0° C. Iodotrimethylsilyl (TMSI) (187 μl, 1.³¹ mmol) was addedand the solution was warmed to rt. After 45 minutes, the solution wascooled again to 0° C. and triethylamine (1 mL, 7.33 mmol) and 2 mL ofMeOH. The solution was warmed to rt and stirred for an additional 20minutes. The solution was concentrated, azeotroped 2× with toluene andput on high vacuum for 30 minutes. Coupling with VII gave compound 37(86.5 mg, 57%) as a yellow solid. ^(1H) NMR (300 MHz, CD₃OD): d 8.30 (d,J=9.5 Hz, 1H), 8.20 (s, 1H), 7.79 (s, 2H), 7.33 (m, 2H), 7.18 (m, 2H),6.98 (t, 1H), 5.95 (m, 1H), 5.78 (s, 1H), 5.22 (d, J=9.6 Hz, 2H), 5.13(d, J=9.0 Hz, 2H), 4.65 (m, 2H), 4.42 (bs, 1H), 4.20 (s, 2H), 4.05 (s,3H), 3.33 (m, 2H), 2.80 (m, 1H), 2.45 (m, 1H), 2.15 (m, 1H), 1.62 (m,6H), 1.38 (d, 6H), 1.05 (s, 9H). ³¹P (121.4 MHz, CD₃OD):

42.855

Example 38 Preparation of Compound 38

The phosphonous acid IV (404 mg, 1.30 mmol) was suspended in 2.5 mL ofCDCl₃. The air was removed from the reaction flask by vacuum andreplaced with N₂. Hunig's Base (543 μl, 3.12 mmol) followed byChlorotrimethylsilyl (363 μl, 2.86 mmol) was added. After 5 minutes,1-(bromomethyl)-4-fluorobenzene (337 μl, 2.77 mmol) was added and thesolution was heated at 40° C. After 4 hours, the reaction wasconcentrated. The residue was partitioned with EtOAc and H₂O and washedwith H₂O. The organic layer was dried over MgSO₄, filtered andconcentrated. The crude material was purified using a CombiFlashChromatography System using a gradient of 50% EtOAc/Hex to 100% EtOAc toobtain(1-benzyloxycarbonylamino-2-vinyl-cyclopropyl)-(4-fluoro-benzyl)-phosphinicacid ethyl ester (164 mg, 30%) as a brown oil. The crude (151 mg, 0.36mmol) was suspended in 1 mL of CH₃CN and cooled to 0° C.Iodotrimethylsilyl (TMSI) (257 μl, 1.81 mmol) was added and the solutionwas warmed to rt. After 45 minutes, the solution was cooled again to 0°C. and triethylamine (1 mL, 7.33 mmol) and 2 mL of MeOH. The solutionwas warmed to rt and stirred for an additional 20 minutes. The solutionwas concentrated, azeotroped 2× with toluene and put on high vacuum for30 minutes. The solid(1-amino-2-vinyl-cyclopropyl)-(4-fluoro-benzyl)-phosphinic acid was useddirectly.

The acid VII (157 mg, 0.24 mmol) was suspended in 1 mL of DMF. HATU (228mg, 0.60 mmol),(1-amino-2-vinyl-cyclopropyl)-(4-fluoro-benzyl)-phosphinic acid (92 mg,0.36 mmol), followed by NMM (132 μl, 1.20 mmol) was added. The solutionstirred overnight at rt. The mixture was purified via Gilson HPLC toobtain 38 (133 mg, 62%) as a yellow solid. ^(1H) NMR (300 MHz, CD₃OD): d8.30 (d, J=9.5 Hz, 1H), 8.20 (s, 1H), 7.79 (s, 2H), 7.35 (d, J=8.8 Hz,1H), 7.05 (t, 2H), 5.95 (m, 1H), 5.78 (s, 1H), 5.35 (d, J=9.6 Hz, 2H),5.15 (d, J=9.0 Hz, 2H), 4.75 (m, 2H), 4.45 (bs, 1H), 4.20 (s, 2H), 4.05(s, 3H), 3.33 (m, 2H), 2.80 (m, 1H), 2.52 (m, 1H), 2.20 (m, 1H), 1.62(m, 6H), 1.38 (d, 6H), 1.05 (s, 9H). ³¹P (121.4 MHz, CD₃OD):

43.659

Example 39 Preparation of Compound 39

A solution of the phosphonous acid IV (330 mg, 1.07 mmol) and Hunig'sBase (392 μl, 2.25 mmol) in CH₂Cl₂ (9.7 mL) stirred at 0° C. aschlorotrimethylsilyl (285 μl, 2.25 mmol) was added dropwise. Thesolution was warmed to rt and after 40 minutes2-(bromomethyl)benzonitrile (461 mg, 2.35 mmol) was added and thesolution was heated at 40° C. for 5 h. The solution stirred at rt for 12h and concentrated. The residue was partitioned with CH₂Cl₂ and NH₄Cl.The organic layer was dried (MgSO₄) and concentrated. The crude materialwas purified with a CombiFlash Chromatography System to give(1-Benzyloxycarbonylamino-2-vinyl-cyclopropyl)-(2-cyano-benzyl)-phosphinicacid ethyl ester (180 mg, 40%). ^(1H) NMR (300 MHz, CDCl₃): d 7.58 (m,4H), 7.33 (m, 5H), 6.18-5.83 (m, 1H), 5.78-5.39 (m, 1H), 5.10 (s, 3H),4.89 (m, 1H), 4.05 (m, 2H), 3.55 (m, 2H), 2.21 (m, 1H), 1.78 (m, 1H),1.50 (m, 1H), 1.10 (m, 3H). ³¹P (121.4 MHz, CDCl₃):

43.997, 41.885 diastereomers

A solution of phosphinate (180 mg, 0.42 mmol) in CH₃CN (1 mL) stirred at0° C. as Iodotrimethylsilyl (301 μl, 2.12 mmol) was added. The solutionstirred at rt then was cooled again to 0° C. and triethylamine (1 mL,7.33 mmol) and MeOH (2 mL) was added. The solution was warmed to rt andstirred for 20 minutes. The solution was concentrated, azeotroped (×2)with toluene and dried on high vacuum for 30 minutes. The solid was usedwithout further purification.

The acid VII (137 mg, 0.21 mmol) was suspended in 3 mL of DMF. HATU (200mg, 0.53 mmol), amine obtained above (111 mg, 0.42 mmol), followed byNMM (136 μl, 1.05 mmol) was added. The solution stirred overnight at rt.The mixture was purified via Gilson HPLC to obtain 39 (43 mg, 23%) as ayellow solid.

^(1H) NMR (300 MHz, CD₃OD): d 8.29 (d, J=8.7 Hz, 1H), 8.18 (s, 1H), 7.76(s, 2H), 7.68 (m, 2H), 7.61 (m, 1H), 7.35 (m, 2H), 5.99 (m, 1H), 5.80(s, 1H), 5.³¹ (d, J=17.5 Hz, 1H), 5.14 (d, J=10.8 Hz, 1H), 4.68 (m, 2H),4.52 (bs, 1H), 4.16 (m, 2H), 4.07 (m, 4H), 3.62 (t, J=15.3 Hz, 1H), 3.42(t, J=15.6 Hz, 1H), 2.83 (m, 1H), 2.66 (m, 1H), 2.18 (m, 1H), 1.62-1.40(m, 10H) 1.38 (d, J=6.3 Hz, 6H), 1.04 (s, 9H). ³¹P (121.4 MHz, CD₃OD):

36.642

LC/MS=898 (M⁺+1)

Example 40 Preparation of Compound 40

The phosphonous acid IV (320 mg, 1.04 mmol) was suspended in 9.7 mL ofCH₂Cl₂. The solution was cooled to 0° C. Hunig's Base (342 μl, 2.25mmol) followed by Chlorotrimethylsilyl (285 μl, 2.25 mmol) was addeddropwise. The solution was warmed to rt and after 40 minutes3-(bromomethyl)benzonitrile (461 mg, 2.35 mmol) was added and thesolution was heated at 40° C. for 5 h. Then the reaction stirred at rtfor 12 h. The residue was partitioned with CH₂Cl₂ and NH₄Cl and washedwith NH₄Cl. The organic layer was dried over MgSO₄, filtered andconcentrated. The crude material was purified using a CombiFlashChromatography System to give(1-Benzyloxycarbonylamino-2-vinyl-cyclopropyl)-(3-cyano-benzyl)-phosphinicacid ethyl ester (190 mg, 42%). ^(1H) NMR (300 MHz, CDCl₃): d 7.58 (m,4H), 7.37 (m, 5H), 6.13 (m, 1H), 5.83-5.78 (m, 1H), 5.65 (m, 1H), 5.39(m, 1H), 5.10 (s, 2H), 3.98 (m, 2H), 3.25 (m, 2H), 2.15 (m, 1H), 1.78(m, 1H), 1.41 (m, 1H), 1.10 (m, 3H). ³¹P (121.4 MHz, CDCl₃):

44.552, 42.103 diastereomers.

Phosphinate (180 mg, 0.42 mmol) was suspended in 1 mL of CH₃CN andcooled to 0° C. Iodotrimethylsilyl(TMSI) (301 μl, 2.12 mmol) was addedand the solution was warmed to rt. After 45 minutes, the solution wascooled again to 0° C. and triethylamine (1 mL, 7.33 mmol) and 2 mL ofMeOH. The solution was warmed to rt and stirred for an additional 20minutes. The solution was concentrated, azeotroped 2× with toluene andput on high vacuum for 30 minutes. The solid was coupled with acid VII(137 mg, 0.21 mmol) in 3 mL of DMF, HATU (200 mg, 0.53 mmol), NMM (136μl, 1.05 mmol). The solution stirred overnight at rt. The mixture waspurified via Gilson HPLC to obtain 40 (40 mg, 22%) as a yellow solid.^(1H) NMR (300 MHz, CD₃OD): d 8.29 (d, J=9.9 Hz, 1H), 8.17 (s, 1H), 7.76(s, 2H), 7.62 (m, 1H), 7.59 (m, 1H), 7.41 (m, 2H), 7.34 (d, J=8.8 Hz,1H), 5.89 (m, 1H), 5.78 (s, 1H), 5.24 (d, J=15.9 Hz, 1H), 5.02 (d,J=10.8 Hz, 1H), 4.66 (m, 2H), 4.46 (bs, 1H), 4.15 (m, 20H), 4.05 (m,4H), 3.22 (m, 2H), 2.78 (m, 1H), 2.49 (m, 1H), 2.09 (m, 1H), 1.62-1.50(m, 10H) 1.34 (d, J=6.3 Hz, 6H), 1.02 (s, 9H). ³¹P (121.4 MHz, CD₃OD):

40.005 LC/MS=898 (M⁺+1)

Example 41 Preparation of Compound 41

The phosphonous acid IV (330 mg, 1.07 mmol) was suspended in 9.7 mL ofCH₂Cl₂. The solution was cooled to 0° C. Hunig's Base (342 μl, 2.25mmol) followed by Chlorotrimethylsilyl (285 μl, 2.25 mmol) was addeddropwise. The solution was warmed to rt and after 40 minutes4-(bromomethyl)benzonitrile (7) (461 mg, 2.35 mmol) was added and thesolution was heated at 40° C. for 5 h. Then the reaction stirred at rtfor 12 h. The residue was partitioned with CH₂Cl₂ and NH₄Cl and washedwith NH₄Cl. The organic layer was dried over MgSO₄, filtered andconcentrated. The crude material was purified with a CombiFlashChromatography System to provide(1-Benzyloxycarbonylamino-2-vinyl-cyclopropyl)-(4-cyano-benzyl)-phosphinicacid ethyl ester (200 mg, 45%). ^(1H) NMR (300 MHz, CDCl₃): d 7.58 (m,4H), 7.37 (m, 5H), 6.13-5.83 (m, 1H), 5.78-5.65 (m, 2H), 5.39 (m, 1H),5.10 (s, 2H), 3.98 (m, 2H), 3.25 (m, 2H), 2.15 (m, 1H), 1.78 (m, 1H),1.41 (m, 1H), 1.10 (m, 3H). ³¹P (121.4 MHz, CDCl₃):

46.164, 43.998 diastereomers.

Phosphinate (180 mg, 0.42 mmol) was suspended in 1 mL of CH₃CN andcooled to 0° C. Iodotrimethylsilyl (TMSI) (301 μl, 2.12 mmol) was addedand the solution was warmed to rt. After 45 minutes, the solution wascooled again to 0° C. and triethylamine (1 mL, 7.33 mmol) and 2 mL ofMeOH. The solution was warmed to rt and stirred for an additional 20minutes. The solution was concentrated, azeotroped 2× with toluene andput on high vacuum for 30 minutes. The solid left was coupled with acidVII (137 mg, 0.21 mmol) in 3 mL of DMF, HATU (200 mg, 0.53 mmol) and NMM(136 μl, 1.05 mmol). The mixture was purified via Gilson HPLC to obtain41 (55 mg, 30%) as a yellow solid. ^(1H) NMR (300 MHz, CD₃OD): d 8.29(d, J=9.3 Hz, 1H), 8.17 (s, 1H), 7.75 (s, 2H), 7.63 (m, 2H), 7.52 (m,2H), 7.32 (m, 1H), 5.89 (m, 1H), 5.79 (s, 1H), 5.26 (d, J=17.1 Hz, 1H),5.06 (d, J=10.8 Hz, 1H), 4.68 (m, 2H), 4.45 (bs, 1H), 4.17 (m, 2H), 4.08(m, 4H), 3.37 (m, 2H), 2.76 (m, 1H), 2.48 (m, 1H), 2.07 (m, 1H),1.61-1.40 (m, 10H) 1.34 (d, J=6.3 Hz, 6H), 1.01 (s, 9H). ³¹P (121.4 MHz,CD₃OD):

36.642

LC/MS=898 (M⁺+1)

Example 42 Preparation of Compound 42

Intermediate IV (2.1 g, 6.79 mmol) was dissolved in THF (20 mL) andcooled to −78° C. A solution of 1 M THF of NaN(TMS)₂ (8.83 mL, 8.83mmol) was added dropwise and the reaction mixture was stirred at −78° C.for 1 h. 2-Methoxybenzyl chloride (1.23 mL, 8.83 mmol) was added and thecold bath was removed. The reaction mixture was stirred at rt for 6 h.The reaction mixture was quenched with NH₄Cl and extracted with EtOAc.The organic layers were washed with brine, dried with Na₂SO₄, filtered,and concentrated. The crude product was purified by combi-flash to give2.15 g of phosphinate in 74% yield.

To a solution of phosphinate obtained above (2.15 g) in TFA (10 mL) atr.t. was added DMS (3 mL) and stirred overnight. The mixture wasconcentrated and co-evaporated with toluene. The residue was dissolvedin 1/1 iPrOH/heptane and washed with 6 N HCl (3×100 mL). The combinedaqueous layers were brought to pH=10 with NaOH in a cold bath. Theaqueous layer was extracted with EtOAc. The organic layers were washedwith brine, dried with Na₂SO₄, and concentrated to give 386 mg of aminewhich was coupled to intermediate VI in DMF and HATU following standardprocedures to give crude product. The crude product was purified bycombi-flash to give 1.1 g of tripeptide in 87% yield.

Tripeptide obtained above (1.1 g, 1.18 mmol) was dissolved in CH₃CN (10mL) and cooled to 0° C. Iodotrimethylsilane (0.85 mL, 5.91 mmol) wasadded dropwise and stirred for 10 minutes. 2,6-Lutidine (0.82 mL) wasadded. MeOH (10 mL) was added and the reaction mixture was concentrated.The crude product was purified by HPLC to give 645 mg of compound 42.

^(1H) NMR (300 MHz, CD₃OD): d 8.30 (d, J=9.5 Hz, 1H), 8.20 (s, 1H), 7.79(s, 2H), 7.35 (d, J=8.8 Hz, 1H), 7.05 (t, 2H), 5.95 (m, 1H), 5.78 (s,1H), 5.35 (d, J=9.6 Hz, 2H), 5.15 (d, J=9.0 Hz, 2H), 4.75 (m, 2H), 4.45(bs, 1H), 4.20 (s, 2H), 4.05 (s, 3H), 3.33 (m, 2H), 2.80 (m, 1H), 2.52(m, 1H), 2.20 (m, 1H), 1.62 (m, 6H), 1.38 (d, 6H), 1.05 (s, 9H). ³¹P(121.4 MHz, CD₃OD):

43.659.

Example 43 Preparation of Compound 43

The phosphonous acid IV (373 mg, 1.21 mmol) was suspended in 5 mL of THFand cooled to −40° C. 1N NaN(TMS)₂ (1.43 mL, 1.43 mmol) was addeddropwise over 15 minutes followed by 1-(chloromethyl)-3-methoxybenzene(212 μl, 1.46 mmol) in 1 mL of THF. The solution stirred from −40° C. tort overnight. The reaction was diluted with EtOAc and quenched with 20mL of 1N HCl. The organic layer was washed with brine, dried over MgSO₄,filtered and concentrated. The crude material was purified using aCombiFlash Chromatography System using a gradient of 30% EtOAc/Hex to100% EtOAc to obtain(1-benzyloxycarbonylamino-2-vinyl-cyclopropyl)-(3-methoxy-benzyl)-phosphinicacid ethyl ester (95.9 mg, 19%) as a brown oil, which was suspended in 1mL of CH₃CN and cooled to 0° C. To this mixture, iodotrimethylsilyl(TMSI) (158 μl, 1.11 mmol) was added and the solution was warmed to rt.After 45 minutes, the solution was cooled again to 0° C. andtriethylamine (1 mL, 7.33 mmol) and 2 mL of MeOH. The solution waswarmed to rt and stirred for an additional 20 minutes. The solution wasconcentrated, azeotroped 2× with toluene and put on high vacuum for 30minutes. The solid(1-Amino-2-vinyl-cyclopropyl)-(3-methoxy-benzyl)-phosphinic acid wascoupled with VII (95 mg, 0.16 mmol), HATU (142 mg, 0.38 mmol) and NMM(83 μl, 0.75 mmol) to give compound 43 after Gilson HPLC purification.^(1H) NMR (300 MHz, CD₃OD): d 8.80 (s, 1H), 8.30 (d, J=9.5 Hz, 1H), 8.20(s, 1H), 7.79 (s, 2H), 7.35 (d, J=8.8 Hz, 1H), 7.18 (m, 1H), 6.85 (m,2H), 6.78 (m, 1H), 5.95 (m, 1H), 5.78 (s, 1H), 5.50 (s, 1H), 5.35 (d,J=9.6 Hz, 2H), 5.15 (d, J=9.0 Hz, 2H), 4.75 (m, 2H), 4.45 (bs, 1H), 4.20(s, 2H), 4.05 (s, 3H), 3.80 (s, 3H), 3.33 (m, 2H), 2.80 (m, 1H), 2.52(m, 1H), 2.20 (m, 1H), 1.62 (m, 6H), 1.38 (d, 6H), 1.05 (s, 9H). ³¹P(121.4 MHz, CD₃OD):

43.371

Example 44 Preparation of Compound 44

The phosphonous acid IV (341 mg, 1.10 mmol) was suspended in 5 mL of THFand cooled to −40° C. 1N NaN(TMS)₂ (1.32 mL, 1.32 mmol) was addeddropwise over 15 minutes followed by 1-(chloromethyl)-4-methoxybenzene(180 μl, 1.32 mmol) in 1 mL of THF. The solution stirred from −40° C. tort overnight. The reaction was diluted with EtOAc and quenched with 20mL of 1N HCl. The organic layer was washed with brine, dried over MgSO₄,filtered and concentrated. The crude material was purified using aCombiFlash Chromatography System using a gradient of 30% EtOAc/Hex to100% EtOAc to obtain(1-Benzyloxycarbonylamino-2-vinyl-cyclopropyl)-(4-methoxy-benzyl)-phosphinicacid ethyl ester (135 mg, 27%) as a brown oil. The residue was suspendedin 1 mL of CH₃CN and cooled to 0° C. Iodotrimethylsilyl (TMSI) (215 μl,1.51 mmol) was added and the solution was warmed to rt. After 45minutes, the solution was cooled again to 0° C. and triethylamine (1 mL,7.33 mmol) and 2 mL of MeOH. The solution was warmed to rt and stirredfor an additional 20 minutes. The solution was concentrated, azeotroped2× with toluene and put on high vacuum for 30 minutes. The solid(1-Amino-2-vinyl-cyclopropyl)-(4-methoxy-benzyl)-phosphinic acid wasused directly.

The acid VII (130 mg, 0.20 mmol) was suspended in 1 mL of DMF. HATU (190mg, 0.50 mmol),(1-Amino-2-vinyl-cyclopropyl)-(4-methoxy-benzyl)-phosphinic acid (80 mg,0.30 mmol), followed by NMM (110 μl, 1.00 mmol) was added. The solutionstirred overnight at rt. The mixture was purified via Gilson HPLC toobtain 44 (85.4 mg, 47%) as a yellow solid. ^(1H) NMR (300 MHz, CD₃OD):d 8.80 (s, 1H), 8.30 (d, J=9.5 Hz, 1H), 8.20 (s, 1H), 7.79 (s, 2H), 7.35(d, J=8.8 Hz, 1H), 7.21 (d, J=8.9 Hz, 2H), 6.85 (d, J=9.2 Hz, 2H), 5.95(m, 1H), 5.78 (s, 1H), 5.50 (s, 1H), 5.35 (d, J=9.6 Hz, 2H), 5.15 (d,J=9.0 Hz, 2H), 4.75 (m, 2H), 4.45 (bs, 1H), 4.20 (s, 2H), 4.05 (s, 3H),3.80 (s, 3H), 3.33 (m, 2H), 2.80 (m, 1H), 2.52 (m, 1H), 2.20 (m, 1H),1.62 (m, 6H), 1.38 (d, 6H), 1.05 (s, 9H). ³¹P (121.4 MHz, CD₃OD):

43.939

Example 45 Preparation of Compound 45

The phosphonous acid IV (330 mg, 1.07 mmol) was suspended in 9.7 mL ofCH₂Cl₂. The solution was cooled to 0° C. Hunig's Base (342 μl, 2.25mmol) followed by Chlorotrimethylsilyl (285 μl, 2.25 mmol) was addeddropwise. The solution was warmed to rt and after 40 minutes1-(bromomethyl)-3-methylbenzene (435 mg, 2.35 mmol) was added and thesolution was heated at 40° C. for 5 h. Then the reaction stirred at rtfor 12 h. The residue was partitioned with CH₂Cl₂ and NH₄Cl and washedwith NH₄Cl. The organic layer was dried over MgSO₄, filtered andconcentrated. The crude material was purified using a CombiFlashChromatography System to give(1-Benzyloxycarbonylamino-2-vinyl-cyclopropyl)-(2-methyl-benzyl)-phosphinicacid ethyl ester (190 mg, 43%). ^(1H) NMR (300 MHz, CDCl₃): d 7.37 (m,5H), 7.18 (m, 4H), 6.13 (m, 1H), 5.78 (m, 1H), 5.39 (m, 1H), 5.10 (m,2H), 3.98 (m, 2H), 3.25 (m, 2H), 2.15 (s, 3H), 1.80 (m, 2H) 1.41 (m,1H), 1.10 (m, 3H). ³¹P (121.4 MHz, CDCl₃):

46.105, 43.225 diastereomers.

Phosphinate (173 mg, 0.42 mmol) was suspended in 1 mL of CH₃CN andcooled to 0° C. Iodotrimethylsilyl (TMSI) (301 μl, 2.12 mmol) was addedand the solution was warmed to rt. After 45 minutes, the solution wascooled again to 0° C. and triethylamine (1 mL, 7.33 mmol) and 2 mL ofMeOH. The solution was warmed to rt and stirred for an additional 20minutes. The solution was concentrated, azeotroped 2× with toluene andput on high vacuum for 30 minutes. The solid was used directly withoutfurther purification.

The acid VII (137 mg, 0.21 mmol) was suspended in 3 mL of DMF. HATU (200mg, 0.53 mmol), crude amine obtained above (105 mg, 0.42 mmol), followedby NMM (136 μl, 1.05 mmol) was added. The solution stirred overnight atrt. The mixture was purified via Gilson HPLC to obtain 45 (60 mg, 34%)as a yellow solid. ^(1H) NMR (300 MHz, CD₃OD): d 8.29 (d, J=9.9 Hz, 1H),8.19 (s, 1H), 7.77 (s, 2H), 7.30 (m, 3H), 7.11 (m, 2H), 5.95 (m, 1H),5.81 (s, 1H), 5.32 (d, J=17.7 Hz, 1H), 5.13 (d, J=10.8 Hz, 1H), 4.67 (m,2H), 4.44 (bs, 1H), 4.16 (m, 2H), 4.08 (m, 4H), 3.30 (m, 2H), 2.75 (m,1H), 2.50 (m, 1H), 2.38 (m, 3H), 2.16 (m, 1H), 1.63-1.35 (m, 6H) 1.34(d, J=6.3 Hz, 6H), 1.03 (s, 9H). ³¹P (121.4 MHz, CD₃OD):

42.100

LC/MS=887 (M⁺+1)

Example 46 Preparation of Compound 46

The phosphonous acid IV (330 mg, 1.07 mmol) was suspended in 9.7 mL ofCH₂Cl₂. The solution was cooled to 0° C. Hunig's Base (342 μl, 2.25mmol) followed by Chlorotrimethylsilyl (285 μl, 2.25 mmol) was addeddropwise. The solution was warmed to rt and after 40 minutes1-(bromomethyl)-3-methylbenzene (435 mg, 2.35 mmol) was added and thesolution was heated at 40° C. for 5 h. Then the reaction stirred at rtfor 12 h. The residue was partitioned with CH₂Cl₂ and NH₄Cl and washedwith NH₄Cl. The organic layer was dried over MgSO₄, filtered andconcentrated. The crude material was purified using a CombiFlashChromatography System to give(1-Benzyloxycarbonylamino-2-vinyl-cyclopropyl)-(3-methyl-benzyl)-phosphinicacid ethyl ester (200 mg, 45%). ^(1H) NMR (300 MHz, CDCl₃): d 7.38 (m,5H), 7.18 (m, 4H), 6.13 (m, 1H), 5.78 (m, 1H), 5.39 (m, 1H), 5.10 (m,2H), 4.02 (m, 2H), 3.25 (m, 2H), 2.30 (s, 3H), 1.98 (m, 1H), 1.80 (m,1H), 1.50 (m, 1H), 1.18 (m, 3H). ³¹P (121.4 MHz, CDCl₃):

7.885, 44.001 diastereomers

Deprotection and coupling as described for example 45 give compound 46as a yellow solid. ^(1H) NMR (300 MHz, CD₃OD): d 8.29 (d, J=9.0 Hz, 1H),8.17 (s, 1H), 7.75 (s, 2H), 7.33 (d, J=9.3, 1H), 7.13 (m, 3H), 7.01 (m,1H), 5.96 (m, 1H), 5.79 (s, 1H), 5.27 (d, J=17.1 Hz, 1H), 5.08 (d, J=9.9Hz, 1H), 4.68 (m, 2H), 4.46 (bs, 1H), 4.16 (m, 2H), 4.06 (m, 4H), 3.27(m, 2H), 2.78 (m, 1H), 2.52 (m, 1H), 2.29 (s, 3H), 2.13 (m, 1H),1.62-1.40 (m, 10H) 1.34 (d, J=6.3 Hz, 6H), 1.03 (s, 9H). ³¹P (121.4 MHz,CD₃OD):

42.100

LC/MS=887 (M⁺+1)

Example 47 Preparation of Compound 47

The phosphonous acid IV (330 mg, 1.07 mmol) was suspended in 9.7 mL ofCH₂Cl₂. The solution was cooled to 0° C. Hunig's Base (342 μl, 2.25mmol) followed by Chlorotrimethylsilyl (285 μl, 2.25 mmol) was addeddropwise. The solution was warmed to rt and after 40 minutes1-(bromomethyl)-4-methylbenzene (435 mg, 2.35 mmol) was added and thesolution was heated at 40° C. for 5 h. Then the reaction stirred at rtfor 12 h. The residue was partitioned with CH₂Cl₂ and NH₄Cl and washedwith NH₄Cl. The organic layer was dried over MgSO₄, filtered andconcentrated. The crude material was purified using a CombiFlashChromatography System to give(1-Benzyloxycarbonylamino-2-vinyl-cyclopropyl)-(4-methyl-benzyl)-phosphinicacid ethyl ester (195 mg, 44%). ^(1H) NMR (300 MHz, CDCl₃): d 7.38 (m,5H), 7.18 (m, 4H), 6.13 (m, 1H), 5.78 (m, 1H), 5.39 (m, 1H), 5.10 (m,2H), 4.04 (m, 2H), 3.25 (m, 2H), 2.30 (s, 3H), 1.98 (m, 1H), 1.80 (m,1H), 1.50 (m, 1H), 1.18 (m, 3H). ³¹P (121.4 MHz, CDCl₃):

45.991, 42.100 diastereomers.

Deprotection and coupling as described for example 45 give compound 47as a yellow solid.

^(1H) NMR (300 MHz, CD₃OD): d 8.26 (d, J=9.5 Hz, 1H), 8.19 (s, 1H), 7.74(s, 2H), 7.30 (m, 1H), 7.19 (m, 2H), 7.06 (m, 2H), 5.94 (m, 1H), 5.78(s, 1H), 5.25 (d, J=17.1 Hz, 1H), 5.06 (d, J=9.0 Hz, 1H), 4.68 (m, 2H),4.47 (bs, 1H), 4.16 (m, 2H), 4.05 (m, 4H), 3.26 (m, 2H), 2.77 (m, 1H),2.48 (m, 1H), 2.27 (s, 3H), 2.09 (m, 1H), 1.65-1.43 (m, 8H) 1.34 (d,J=6.3 Hz, 6H), 1.02 (s, 9H). ³¹P (121.4 MHz, CD₃OD):

42.100 LC/MS=887 (M⁺+1)

Example 48 Preparation of Compound 48

The phosphonous acid IV (330 mg, 1.07 mmol) was suspended in 9.7 mL ofCH₂Cl₂. The solution was cooled to 0° C. Hunig's Base (342 μl, 2.25mmol) followed by Chlorotrimethylsilyl (285 μl, 2.25 mmol) was addeddropwise. The solution was warmed to rt and after 40 minutes1-(bromomethyl)-1-(trifluoromethyl)benzene (456 mg, 2.35 mmol) was addedand the solution was heated at 40° C. for 48 h. Then the reactionstirred at rt for 12 h. The residue was partitioned with CH₂Cl₂ andNH₄Cl and washed with NH₄Cl. The organic layer was dried over MgSO₄,filtered and concentrated. The crude material was purified using aCombiFlash Chromatography System give(1-Benzyloxycarbonylamino-2-vinyl-cyclopropyl)-(2-trifluoromethyl-benzyl)-phosphinicacid ethyl ester (225 mg, 45%). ^(1H) NMR (300 MHz, CDCl₃): d 7.61 (m,2H), 7.43 (m, 2H), 7.33 (m, 5H), 6.13 (m, 1H), 5.83 (m, 1H), 5.78 (m,1H), 5.39 (m, 1H), 5.10 (s, 2H), 3.88 (m, 2H), 3.25 (m, 2H), 2.10 (m,1H), 1.78 (m, 1H), 1.41 (m, 1H), 1.10 (m, 3H). ³¹P (121.4 MHz, CDCl₃):

45.337, 42.005 diastereomers.

Deprotection and coupling as described for example 45 give compound 48as a yellow solid (80 mg, 45%).

¹H NMR (300 MHz, CD₃OD): d 8.28 (d, J=9.3 Hz, 1H), 8.19 (s, 1H), 7.76(s, 2H), 7.69 (m, 2H), 7.53 (t, J=7.8 Hz, 1H), 7.39 (t, J=7.5 Hz, 1H),7.32 (d, J=9.3 Hz, 1H), 5.95 (m, 1H), 5.82 (s, 1H), 5.30 (d, J=17.4 Hz,1H), 5.12 (d, J=11.1 Hz, 1H), 4.69 (m, 2H), 4.43 (bs, 1H), 4.17 (m, 2H),4.06 (m, 4H), 3.65 (t, J=15.3 Hz, 1H), 3.43 (t, J=16.5 Hz, 1H), 2.79 (m,1H), 2.53 (m, 1H), 2.17 (m, 1H), 1.70-1.40 (m, 10H), 1.34 (d, J=6.3 Hz,6H), 1.01 (s, 9H).

³¹P (121.4 MHz, CD₃OD):

40.995. LC/MS=941 (M⁺+1)

Example 49 Preparation of Compound 49

The phosphonous acid IV (330 mg, 1.07 mmol) was suspended in 9.7 mL ofCH₂Cl₂. The solution was cooled to 0° C. Hunig's Base (342 μl, 2.25mmol) followed by Chlorotrimethylsilyl (285 μl, 2.25 mmol) was addeddropwise. The solution was warmed to rt and after 40 minutes1-(bromomethyl)-3-(trifluoromethyl)benzene (456 mg, 2.35 mmol) was addedand the solution was heated at 40° C. for 48 h. Then the reactionstirred at rt for 12 h. The residue was partitioned with CH₂Cl₂ andNH₄Cl and washed with NH₄Cl. The organic layer was dried over MgSO₄,filtered and concentrated. The crude material was purified using aCombiFlash Chromatography System give(1-Benzyloxycarbonylamino-2-vinyl-cyclopropyl)-(3-trifluoromethyl-benzyl)-phosphinicacid ethyl ester (230 mg, 46%). ^(1H) NMR (300 MHz, CDCl₃): d 7.43 (m,4H), 7.33 (m, 5H), 6.13 (m, 1H), 5.83 (m, 1H), 5.78 (m, 1H), 5.39 (m,1H), 5.10 (s, 3H), 3.88 (m, 2H), 3.25 (m, 2H), 2.11 (m, 1H), 1.78 (m,1H), 1.41 (m, 1H), 1.10 (m, 3H). ³¹P (121.4 MHz, CDCl₃):

45.337, 42.005 diastereomers.

Deprotection and coupling as described for example 45 give compound 49as a yellow solid (80 mg, 45%).

^(1H) NMR (300 MHz, CD₃OD): d 8.28 (d, J=9.3 Hz, 1H), 8.18 (s, 1H), 7.75(s, 2H), 7.65 (m, 1H), 7.59 (m, 1H), 7.46 (m, 2H), 7.³¹ (d, J=9.0 Hz,1H), 5.87 (m, 1H), 5.80 (s, 1H), 5.23 (d, J=17.1 Hz, 1H), 5.02 (d,J=10.2 Hz, 1H), 4.66 (m, 2H), 4.46 (bs, 1H), 4.18 (m, 2H), 4.07 (m, 4H),3.38 (m, 2H), 2.81 (m, 1H), 2.51 (m, 3H), 2.10 (m, 1H), 1.63-1.50 (m,10H), 1.34 (d, J=6.3 Hz, 6H), 1.05 (s, 9H). ³¹P (121.4 MHz, CD₃OD):

40.995 LC/MS=941 (M⁺+1)

Example 50 Preparation of Compound 50

The phosphonous acid IV (330 mg, 1.07 mmol) was suspended in 9.7 mL ofCH₂Cl₂. The solution was cooled to 0° C. Hunig's Base (342 μl, 2.25mmol) followed by Chlorotrimethylsilyl (285 μl, 2.25 mmol) was addeddropwise. The solution was warmed to rt and after 40 minutes1-(bromomethyl)-4-(trifluoromethyl)benzene (28) (456 mg, 2.35 mmol) wasadded and the solution was heated at 40° C. for 48 h. Then the reactionstirred at rt for 12 h. The residue was partitioned with CH₂Cl₂ andNH₄Cl and washed with NH₄Cl. The organic layer was dried over MgSO₄,filtered and concentrated. The crude material was purified using aCombiFlash Chromatography System give(1-Benzyloxycarbonylamino-2-vinyl-cyclopropyl)-(4-trifluoromethyl-benzyl)-phosphinicacid ethyl ester (205 mg, 41%). ^(1H) NMR (300 MHz, CDCl₃): d 7.55 (m,4H), 7.39 (m, 5H), 6.13 (m, 1H), 5.83 (m, 1H), 5.78 (m, 1H), 5.39 (m,1H), 5.10 (s, 2H), 3.88 (m, 2H), 3.25 (m, 2H), 2.03 (m, 1H), 1.78 (m,1H), 1.41 (m, 1H), 1.10 (m, 3H). ³¹P (121.4 MHz, CDCl₃):

45.337, 42.005 diastereomers.

Deprotection and coupling as described for example 45 give compound 50as a yellow solid (80 mg, 45%).

^(1H) NMR (300 MHz, CD₃OD): d 8.27 (d, J=9.3 Hz, 1H), 8.19 (s, 1H), 7.75(s, 2H), 7.51 (m, 4H), 7.27 (m, 1H), 5.92 (m, 1H), 5.81 (s, 1H), 5.23(d, J=17.1 Hz, 1H), 5.04 (d, J=10.5 Hz, 1H), 4.67 (m, 2H), 4.46 (bs,1H), 4.17 (m, 2H), 4.04 (m, 4H), 3.35 (m, 2H), 2.80 (m, 1H), 2.49 (m,1H), 2.08 (m, 1H), 1.62-1.39 (m, 8H), 1.32 (d, J=6.3, 6H), 1.02 (s, 9H).³¹P (121.4 MHz, CD₃OD):

40.995

LC/MS=941 (M⁺+1)

Example 51 Preparation of Compound 51

Intermediate IV (13.42 g, 43.4 mmol) was dissolved in CH₂Cl₂ (300 mL)and cooled to 0° C. and diisopropylethylamine (15.4 mL, 91.1 mmol) wasadded. Chlorotrimethylsilane (11.4 mL, 91.1 mmol) was added dropwise.The reaction mixture was warmed to r.t. and stirred for 1.5 h.2-chlorobenzyl chloride (15.6 g, 95.5 mmol) was added and the reactionwas heated to 50° C. for 48 h. The reaction mixture was cooled to rt,diluted with CH₂Cl₂, washed with aqueous NH₄Cl, dried with Na₂SO₄, andconcentrated. The crude product was purified by combi-flash to givephosphinate.

The phosphinate (5.0 g, 11.55 mmol) (147 mg, 0.34 mmol) was suspended in1 mL of CH₃CN and cooled to 0° C. Iodotrimethylsilyl (TMSI) (241 μl,1.70 mmol) was added and the solution was warmed to rt. After 45minutes, the solution was cooled again to 0° C. and triethylamine (1 mL,7.33 mmol) and 2 mL of MeOH. The solution was warmed to rt and stirredfor an additional 20 minutes. The solution was concentrated, azeotroped2× with toluene and put on high vacuum for 30 minutes. The crude wascoupled to acid VII to give compound 51. ^(1H) NMR (300 MHz, CD₃OD): d8.30 (d, J=9.5 Hz, 1H), 8.20 (s, 1H), 7.79 (s, 2H), 7.50 (d, J=8.8 Hz,1H), 7.38 (m, 2H), 7.20 (m, 2H), 5.95 (m, 1H), 5.80 (s, 1H), 5.25 (d,J=9.6 Hz, 2H), 5.15 (d, J=9.0 Hz, 2H), 4.75 (m, 2H), 4.45 (bs, 1H), 4.20(s, 2H), 4.05 (s, 3H), 3.33 (s, 2H), 2.80 (m, 1H), 2.52 (m, 1H), 2.15(m, 1H), 1.62 (m, 6H), 1.38 (d, 6H), 1.05 (s, 9H). ³¹P (121.4 MHz,CD₃OD):

42.155

Example 52 Preparation of Compound 52

The phosphonous acid IV (369 mg, 1.19 mmol) was suspended in 5 mL of THFand cooled to −40° C. 1N NaN(TMS)₂ (1.43 mL, 1.43 mmol) was addeddropwise over 15 minutes followed by 1-chloro-3-(chloromethyl)benzene(182 μl, 1.43 mmol) in 1 mL of THF. The solution stirred from −40° C. tort overnight. The reaction was diluted with EtOAc and quenched with 20mL of 1N HCl. The organic layer was washed with brine, dried over MgSO₄,filtered and concentrated. The crude material was purified using aCombiFlash Chromatography System using a gradient of 30% EtOAc/Hex to100% EtOAc to obtain(1-benzyloxycarbonylamino-2-vinyl-cyclopropyl)-(3-chloro-benzyl)-phosphinicacid ethyl ester (90.5 mg, 24%) as a brown oil. The crude was suspendedin 1 mL of CH₃CN and cooled to 0° C. Iodotrimethylsilyl (TMSI) (148 μl,1.04 mmol) was added and the solution was warmed to rt. After 45minutes, the solution was cooled again to 0° C. and triethylamine (1 mL,7.33 mmol) and 2 mL of MeOH. The solution was warmed to rt and stirredfor an additional 20 minutes. The solution was concentrated, azeotroped2× with toluene and put on high vacuum for 30 minutes. The solid(1-Amino-2-vinyl-cyclopropyl)-(3-chloro-benzyl)-phosphinic acid was useddirectly.

The acid (87 mg, 0.13 mmol) was suspended in 1 mL of DMF. HATU (123 mg,0.33 mmol), VII (54.6 mg, 0.20 mmol), followed by NMM (71 μl, 0.65 mmol)was added. The solution stirred overnight at rt. The mixture waspurified via Gilson HPLC to obtain 52 (68.5 mg, 59%) as a yellow solid.^(1H) NMR (300 MHz, CD₃OD): d 8.30 (d, J=9.5 Hz, 1H), 8.20 (s, 1H), 7.79(s, 2H), 7.40 (s, 1H), 7.35 (d, J=8.8 Hz, 1H), 7.23 (m, 1H), 6.85 (m,2H), 6.78 (m, 1H), 5.95 (m, 1H), 5.78 (s, 1H), 5.50 (s, 1H), 5.35 (d,J=9.6 Hz, 2H), 5.15 (d, J=9.0 Hz, 2H), 4.75 (m, 2H), 4.45 (bs, 1H), 4.20(s, 2H), 4.05 (s, 3H), 3.33 (m, 2H), 2.80 (m, 1H), 2.52 (m, 1H), 2.20(m, 1H), 1.62 (m, 6H), 1.38 (d, 6H), 1.05 (s, 9H). ³¹P (121.4 MHz,CD₃OD):

42.73

Example 53 Preparation of Compound 53

The phosphonous acid IV (370 mg, 1.20 mmol) was suspended in 5 mL of THFand cooled to −40° C. 1N NaN(TMS)₂ (1.43 mL, 1.43 mmol) was addeddropwise over 15 minutes followed by 1-chloro-4-(chloromethyl)benzene(2³¹ mgl, 1.43 mmol) in 1 mL of THF. The solution stirred from −40° C.to rt overnight. The reaction was diluted with EtOAc and quenched with20 mL of 1N HC. The organic layer was washed with brine, dried overMgSO₄, filtered and concentrated. The crude material was purified usinga CombiFlash Chromatography System using a gradient of 30% EtOAc/Hex to100% EtOAc to obtain(1-Benzyloxycarbonylamino-2-vinyl-cyclopropyl)-(4-chloro-benzyl)-phosphinicacid ethyl ester (94 mg, 26%) as a brown oil. The residue (94.9 mg, 0.22mmol) was suspended in 1 mL of CH₃CN and cooled to 0° C.Iodotrimethylsilyl (TMSI) (155 μl, 1.09 mmol) was added and the solutionwas warmed to rt. After 45 minutes, the solution was cooled again to 0°C. and triethylamine (1 mL, 7.33 mmol) and 2 mL of MeOH. The solutionwas warmed to rt and stirred for an additional 20 minutes. The solutionwas concentrated, azeotroped 2× with toluene and put on high vacuum for30 minutes. The phosphinic acid was coupled with intermediate VII (96mg, 0.15 mmol) in 1 mL of DMF, HATU (142 mg, 0.37 mmol), and NMM (821,0.75 mmol) to give 53 (75.2 mg, 55%) as a yellow solid. ^(1H) NMR (300MHz, CD₃OD): d 8.30 (d, J=9.5 Hz, 1H), 8.20 (s, 1H), 7.79 (s, 2H), 7.30(m, 4H), 5.95 (m, 1H), 5.80 (s, 1H), 5.25 (d, J=9.6 Hz, 2H), 5.13 (d,J=9.0 Hz, 2H), 4.75 (m, 2H), 4.45 (bs, 1H), 4.20 (s, 2H), 4.05 (s, 3H),3.33 (s, 2H), 2.80 (m, 1H), 2.52 (m, 1H), 2.15 (m, 1H), 1.62 (m, 6H),1.38 (d, 6H), 1.05 (s, 9H). ³¹P (121.4 MHz, CD₃OD):

42.837

Example 54 Preparation of Compound 54

Intermediate IV (398 mg, 1.3 mmol) was dissolved in CH₂Cl₂ (25 mL) andcooled to 0° C. Diisopropylethylamine (0.5 mL, 2.7 mmol) was added andstirred for 25 minutes. Chlorotrimethylsilane (0.4 mL, 2.7 mmol) wasadded dropwise. The reaction mixture was warmed to r.t. and stirred for1 h. 2-Bromobenzyl bromide (1.6 g, 6.4 mmol) was added and the reactionwas heated to 45° C. for 18 h. The reaction mixture was cooled to rt,diluted with CH₂Cl₂, washed with aqueous NH₄Cl, dried with Na₂SO₄, andconcentrated. The crude product was purified by combi-flash to give 346mg of phosphinate in 56% yield.

To a solution of phosphinate obtained above (346 mg, 0.72 mmol) in CH₃CN(1 mL) at 0° C. was added iodotrimethylsilane (0.6 mL, 3.6 mmol). Thereaction mixture was warmed to r.t. and stirred for 30 minutes and thencooled to 0° C. 2,6-Lutidine (0.5 mL) and MeOH (1 mL) were added andstirred for 10 minutes. The solvent was concentrated and the residue wasco-evaporated with toluene (5 mL), and dried under vacuum for 20 minutesto give crude amine. Coupling with acid VII (230 mg, 0.36 mmol) provided360 mg of compound 54. ¹H NMR (300 MHz, DMSO):

8.30 (s, 1H), 8.20 (s, 1H), 7.88 (s, 1H), 7.79 (s, 1H), 7.54 (d, J=7.5Hz, 1H), 7.45 (d, J=7.8 Hz, 1H), 7.29 (m, 1H), 7.11 (dd, J=7.8, 7.8 Hz,1H), 7.01 (d, J=7.5 Hz, 1H), 6.00 (m, 1H), 5.78 (s, 1H), 5.15 (d, J=17.1Hz, 1H), 5.00 (d, J=11.7 Hz, 1H), 4.60-4.40 (m, 3H), 3.43 (dd, J=15.1,15.1 Hz, 1H), 3.25 (dd, J=15.9, 15.9 Hz, 2H), 2.58 (m, 1H), 2.32 (m,1H), 1.94 (m, 1H), 1.70-1.40 (m, 8H), 1.29 (m, 6H), 0.92 (s, 9H). ³¹P(121.4 MHz, CDCl₃) d 39.975. LC/MS=951.20 (M⁺+1), 975.20 (M⁺+Na)

Example 55 Preparation of Compound 55

^(1H) NMR (300 MHz, CD₃OD): d 8.28 (d, J=9.3 Hz, 1H), 8.17 (s, 1H), 7.76(s, 2H), 7.56 (m, 1H), 7.33 (m, 4H), 5.96 (m, 1H), 5.81 (bs, 1H), 5.³¹(d, J=17.1 Hz, 1H), 5.12 (d, J=10.5 Hz, 1H), 4.69 (m, 2H), 4.45 (bs,1H), 4.18 (m, 2H), 4.06 (m, 4H), 3.40 (m, 2H), 2.80 (m, 1H), 2.52 (m,1H), 2.16 (m, 1H), 1.68-1.50 (m, 10H) 1.34 (d, J=6.3 Hz, 6H), 1.01 (s,9H). ³¹P (121.4 MHz, CD₃OD):

40.042. LC/MS=957 (M⁺+1)

Example 56 Preparation of Compound 56

The phosphonous acid IV (1.5 g, 4.85 mmol) was suspended in 40 mL ofCH₂Cl₂. The solution was cooled to 0° C. Hunig's Base (1.73 mL, 10.2mmol) followed by Chlorotrimethylsilyl (1.28 mL, 10.2 mmol) was addeddropwise. The solution was warmed to rt and after 40 minutes1-Bromomethyl-2-isopropoxy-benzene (2.45 g, 10.7 mmol) was added and thesolution was heated at 40° C. for 12 hours. Then the reaction stirred atrt for 12 hours. The residue was partitioned with CH₂Cl₂ and NH₄Cl andwashed with NH₄Cl. The organic layer was dried over MgSO₄, filtered andconcentrated. The crude material was purified using a CombiFlashChromatography System to give(1-Benzyloxycarbonylamino-2-vinyl-cyclopropyl)-(2-isopropoxy-benzyl)-phosphinicacid ethyl ester (1.1 g, 50%). ^(1H) NMR (300 MHz, CDCl₃): d 7.33 (m,5H), 7.10 (m, 2H), 6.89 (m, 2H), 6.18-5.83 (m, 1H), 5.78-5.39 (m, 1H),5.10 (m, 3H), 4.89 (m, 1H), 4.05 (m, 2H), 3.55 (m, 2H), 2.97 (m, 1H),2.01 (m, 1H), 1.78 (m, 1H), 1.50 (m, 1H), 1.20 (m, 9H). ³¹P (121.4 MHz,CDCl₃):

45.097, 44.785 diastereomers.

The phosphinate (700 mg, 1.07 mmol) was suspended in 1 mL of CH₃CN andcooled to 0° C. Iodotrimethylsilyl (TMSI) (727 μl, 5.35 mmol) was addedand the solution was warmed to rt. After 45 minutes, the solution wascooled again to 0° C. and triethylamine (2 mL, 14.6 mmol) and 2 mL ofMeOH. The solution was warmed to rt and stirred for an additional 20minutes. The solution was concentrated, azeotroped 2× with toluene andput on high vacuum for 30 minutes. The solid amine was coupled to acidVII to provide compound 56. ^(1H) NMR (300 MHz, CD₃OD): d 8.28 (d, J=9.6Hz, 1H), 8.18 (s, 1H), 7.75 (s, 2H), 7.38 (m, 2H), 7.17 (m, 1H), 6.92(m, 1H), 6.82 (m, 1H), 5.95 (m, 1H), 5.80 (s, 1H), 5.27 (d, J=17.1 Hz,1H), 5.06 (d, J=9.0 Hz, 1H), 4.63 (m, 4H), 4.46 (bs, 1H), 4.17 (m, 2H),4.07 (m, 4H), 3.34 (m, 3H), 2.73 (m, 1H), 2.51 (m, 1H), 2.13 (m, 1H),1.62 (m, 1H), 1.50 (m, 8H) 1.38 (m, 12H), 1.05 (s, 9H). ³¹P (121.4 MHz,CD₃OD):

36.642. LC/MS=9³¹ (M⁺+1)

Example 57 Preparation of Compound 57

To a solution of (2-ethyl-phenyl)-methanol (3 g, 22 mmol) in ether (10mL) at 0° C. was added a solution of PBr₃ (2.18 g, 8.1 mmol) in ether (3mL). The reaction mixture was warmed to r.t for 45 minutes and cooled to0° C. The reaction mixture was treated with 50% aqueous KOH (15 mL) andseparated. The organic layer was dried with KOH pellets and concentratedto give 3.9 g of 1-bromomethyl-2-ethyl-benzene.

Intermediate IV (1.29 g, 3.88 mmol) was dissolved in CH₂Cl₂ (40 mL) andcooled to 0° C. Diisopropylethylamine (1.41 mL, 8.15 mmol) was added andstirred for 15 minutes. Chlorotrimethylsilane (1.1 mL, 8.15 mmol) wasadded dropwise. The reaction mixture was warmed to r.t. and stirred for1 h. 2-ethylbenzyl bromide (3.86 g, 19.4 mmol) was added and thereaction was heated to 45° C. overnight. The reaction mixture was cooledto rt, diluted with CH₂Cl₂, washed with aqueous NH₄Cl, dried withNa₂SO₄, and concentrated. The crude product was purified by combi-flashto give 683 mg of phosphinate in 41% yield.

To a solution of phosphinate obtained above (650 mg, 1.52 mmol) in CH₃CN(3 mL) at 0° C. was added iodotrimethylsilane (1.52 g, 7.6 mmol). Thereaction mixture was warmed to r.t. and stirred for 30 minutes and thencooled to 0° C. 2,6-Lutidine (0.9 mL) and MeOH (1.5 mL) were added andstirred for 10 minutes. The solvent was concentrated and the residue wasco-evaporated with toluene (5 mL), and dried under vacuum for 20 minutesto give crude amine which was coupled to VII (500 mg, 0.76 mmol) to givecompound 57 (480 mg, 70%). ¹H NMR (300 MHz, DMSO): d 8.28 (s, 1H), 8.21(d, J=9.6 Hz, 1H), 7.88 (s, 1H), 7.79 (s, 1H), 7.32 (d, J=11.1 Hz, 1H),7.19-7.01 (m, 4H), 6.00 (m, 1H), 5.78 (s, 1H), 5.17 (d, J=17.1 Hz, 1H),5.02 (d, J=12.3 Hz, 1H), 4.54 (m, 2H), 4.47 (bs, 1H), 4.16 (m, 3H), 3.97(s, 3H), 3.15 (m, 2H), 2.60 (m, 1H), 2.29 (m, 3H), 1.94 (m, 1H),1.70-2.40 (m, 8H), 1.30 (m, 6H), 0.92 (s, 9H). ³¹P (121.4 MHz, CDCl₃): d40.942. LC/MS=901.24 (M⁺+1), 924.17 (M⁺+Na)

Example 58 Preparation of Compound 58

The phosphonous acid IV (327 mg, 1.06 mmol) was suspended in 5 mL of THFand cooled to −40° C. 1N NaN(TMS)₂ (1.27 mL, 1.39 mmol) was addeddropwise over 15 minutes followed by 2-(bromomethyl)-1,3-difluorobenzene(176 μl, 1.39 mmol) in 1 mL of THF. The solution stirred from −40° C. tort overnight. The reaction was diluted with EtOAc and quenched with 20mL of 1N HCl. The organic layer was washed with brine, dried over MgSO₄,filtered and concentrated. The crude material was purified using aCombiFlash Chromatography System using a gradient of 30% EtOAc/Hex to100% EtOAc to obtain(1-benzyloxycarbonylamino-2-vinyl-cyclopropyl)-(2,6-difluoro-benzyl)-phosphinicacid ethyl ester (147 mg, 33%) as a brown oil. The phosphinate (94.7 mg,0.22 mmol) was suspended in 1 mL of CH₃CN and cooled to 0° C.Iodotrimethylsilyl (TMSI) (155 μl, 1.08 mmol) was added and the solutionwas warmed to rt. After 45 minutes, the solution was cooled again to 0°C. and triethylamine (1 mL, 7.33 mmol) and 2 mL of MeOH. The solutionwas warmed to rt and stirred for an additional 20 minutes. The solutionwas concentrated, azeotroped 2× with toluene and put on high vacuum for30 minutes to provide crude amine,(1-Amino-2-vinyl-cyclopropyl)-(2,6-difluoro-benzyl)-phosphinic acidethyl ester.

The acid VII (96 mg, 0.15 mmol) was suspended in 1 mL of DMF. HATU (143mg, 0.37 mmol), amine obtained above (60 mg, 0.22 mmol) was added,followed by the addition of NMM (83 μl, 0.75 mmol). The solution wasstirred overnight at rt. The mixture was purified via Gilson HPLC toobtain 58 (67 mg, 53%) as a yellow solid. ¹H NMR (300 MHz, CD₃OD): d8.30 (d, J=9.5 Hz, 1H), 8.20 (s, 1H), 7.79 (s, 2H), 7.35 (d, J=9.3 Hz,1H), 7.33 (m, 1H), 6.94 (m, 2H), 5.95 (m, 1H), 5.80 (s, 1H), 5.25 (d,J=9.6 Hz, 2H), 5.17 (d, J=9.0 Hz, 2H), 4.75 (m, 2H), 4.45 (bs, 1H), 4.20(s, 2H), 4.05 (s, 3H), 3.40 (m, 2H), 2.80 (m, 1H), 2.52 (m, 1H), 2.15(m, 1H), 1.62 (m, 6H), 1.38 (d, 6H), 1.05 (s, 9H). ³¹P (121.4 MHz,CD₃OD):

40.898

Example 59 Preparation of Compound 59

Boc-hydroxyproline methyl ester (5 g, 20.4 mmol) was taken up in DCM (50mL) and TFA (50 mL). The reaction was stirred at room temp for 1.5 hthen concentrated and azeotroped with toluene (2×50 mL). The residue wastaken up in DCM (200 mL) and the cyclopentyl carbamate of tert-leucine(5.5 g, 22.4 mmol) was added, followed by HATU (11.6 g, 30.6 mmol) andNMM (9 mL, 81.6 mmol). The reaction was stirred at room temp overnight,then quenched with sat'd NH₄Cl solution, washed with water then brine,dried, and concentrated. The residue was then purified by flashchromatography to provide the desired dipeptide (7.56 g).

The methyl ester was taken up in THF (70 mL), water (70 mL), methanol(70 mL) and LiOH—H₂O (8.6 g, 204 mmol) was added. The reaction wasstirred at room temp for 1 h, then diluted with water and acidified withHCl. The reaction was extracted with ethyl acetate, washed with brine,dried and concentrated to provide the desired acid (5.98 g crude, 82%two steps).

The carboxylic acid (2.62 g, 7.36 mmol) was taken up in THF (75 mL) at0° C. and TEA (3.1 mL, 22.08 mmol) and ethyl chloroformate (0.70 mL,7.36 mmol) were added. The reaction was allowed to warm to room temp andstirred 30 minutes. The solids were filtered off and the reaction wasconcentrated. The residue was taken up in ethyl acetate, washed with 1NHCl, concentrated and purified via flash chromatography to provide thedesired lactone (1.81 g, 73%).

This lactone (0.44 g, 1.3 mmol) was taken up in toluene (8 mL) and water(8 mL) in the presence of the amine prepared in example 83 (0.25 g, 0.83mmol). Sodium ethylhexanoate (0.32 g, 1.95 mmol) was added and thereaction stirred at 80° C. overnight. The reaction was extracted withethyl acetate, washed with sodium bicarbonate solution, 1N HCl, andbrine, dried, concentrated, and purified by flash column to provide thetripeptide (0.25 g, 50%).

The prolinol (0.93 g, 1.45 mmol) was combined with brosyl chloride (0.52g, 2.03 mmol) and DABCO (0.26 g, 2.32 mmol) in toluene (3 mL) andstirred at room temp for 3 h. The reaction was extracted with ethylacetate, washed with sodium bicarbonate solution, 1N HCl, brine,concentrated and purified by flash chromatography to provide thebrosylate (0.995 g, 80%).

The brosylate (0.995 g, 1.16 mmol) was taken up in NMP (12 mL) and8-chloro-4-hydroxy-7-methoxy-quinoline-2-carboxylic acid (0.38 g, 1.16mmol) and cesium carbonate (0.38 g, 1.16 mmol) were added. The reactionwas stirred at 60° C. for 4 hours then room temp overnight. The reactionwas extracted with ethyl acetate, washed with bicarbonate solution,concentrated and purified by flash chromatography to provide the product(0.86 g, 84%).

This methyl ester (0.86 g, 0.97 mmol) was taken up in THF (10 mL) andwater (10 mL) and NaOH (2 mL of 1M solution) were added at 0° C. Thereaction was stirred for 1.5 h, diluted with water, acidified with HCland extracted with ethyl acetate. The organics were dried andconcentrated to provide the carboxylic acid. This residue was taken upin THF and TEA (0.15 mL, 1.07 mmol) was added and the mixture cooled tozero. Isobutylchloroformate (0.14 mL, 1.07 mmol) was added and thereaction stirred at room temp for 40 minutes. Diazomethane (2.0equivalents) was added in ether solution (prepared from MNNG) and thereaction stirred at zero for 30 minutes then for 2 h at room temp. Thereaction was then concentrated to provide the diazoketone (0.58 g, 67%two steps).

The diazoketone (0.58 g, 0.646 mmol) was taken up in THF at 0° C. andcone HBr (0.4 mL) was added. The reaction was stirred and monitored byLCMS. Upon full conversion ethyl acetate was added and the mixture waswashed with NaHCO₃ solution, dried and concentrated. The residue wastaken up in IPA (10 mL) and isopropylthiourea (0.15 g, 1.29 mmol) wasadded. The reaction was heated to 75° C. for 1 h, then concentrated. Theresultant residue was taken up in acetonitrile and TMSI (0.5 mL, 3.23mmol) was added. The reaction was stirred at room temp for 15 minutes,diluted with 0.4 mL of 2,6-lutidine, then quenched with methanol,concentrated and purified by HPLC to provide Compound 59 (443 mg, 73%).¹H NMR (300 MHz, CD₃OD) d 8.29 (m, 2H), 7.79 (m, 1H), 7.59 (m, 1H), 7.24(m, 1H), 6.86 (m, 2H), 5.97 (m, 1H), 5.75 (br s, 1H), 5.32 (m, 1H), 5.09(m, 1H), 4.77 (m, 1H), 4.60 (m, 1H), 4.40 (br s, 1H), 4.14 (s, 3H), 3.95(m, 1H), 3.41 (m, 3H), 2.73 (m, 2H), 2.18 (m, 1H), 1.61 (m, 8H), 1.47(m, 8H), 1.03 (s, 9H). ³¹P NMR (121.4 MHz, CD₃OD)

40.279. LCMS: 943 (M⁺+1).

Example 60 Preparation of Compound 60

The phosphonous acid IV (1.0 g, 3.24 mmol) was suspended in 30 mL ofCH₂Cl₂. The solution was cooled to 0° C. Hunig's Base (1036 μl, 6.79mmol) followed by Chlorotrimethylsilyl (863 μl, 6.79 mmol) was addeddropwise. The solution was warmed to rt and after 40 minutes2-Bromomethyl-1-chloro-3-fluoro-benzene (1.58 g, 7.13 mmol) was addedand the solution was heated at 40° C. for 12 hours. Then the reactionstirred at rt for 12 hours. The residue was partitioned with CH₂Cl₂ andNH₄Cl and washed with NH₄Cl. The organic layer was dried over MgSO₄,filtered and concentrated. The crude material was purified using aCombiFlash Chromatography System to give(1-Benzyloxycarbonylamino-2-vinyl-cyclopropyl)-(2-chloro-6-fluoro-benzyl)-phosphinicacid ethyl ester (750 mg, 51%). ^(1H) NMR (300 MHz, CDCl₃): d 7.33 (m,5H), 7.17 (m, 2H), 6.95 (m, 1H), 6.18-5.83 (m, 1H), 5.78-5.39 (m, 1H),5.10 (m, 3H), 4.89 (m, 1H), 4.05 (m, 2H), 3.55 (m, 2H), 2.21 (m, 1H),1.78 (m, 1H), 1.50 (m, 1H), 1.10 (m, 3H). ³¹P (121.4 MHz, CDCl₃):

45.897, 42.185 diastereomers.

The phosphinate obtained above (730 mg, 1.62 mmol) was suspended in 1 mLof CH₃CN and cooled to 0° C. Iodotrimethylsilyl (TMSI) (1112 μl, 8.18mmol) was added and the solution was warmed to rt. After 45 minutes, thesolution was cooled again to 0° C. and triethylamine (2 mL, 14.6 mmol)and 2 mL of MeOH. The solution was warmed to rt and stirred for anadditional 20 minutes. The solution was concentrated, azeotroped 2× withtoluene and put on high vacuum for 30 minutes. The crude amine was useddirectly. Coupling with VII gave compound 60. ^(1H) NMR (300 MHz,CD₃OD): d 8.23 (d, J=9.5 Hz, 1H), 8.20 (s, 1H), 7.79 (s, 2H), 7.33 (m,1H), 7.21 (m, 2H), 7.03 (m, 1H), 5.95 (m, 1H), 5.78 (s, 1H), 5.22 (d,J=9.6 Hz, 1H), 5.13 (d, J=9.0 Hz, 1H), 4.63 (m, 2H), 4.45 (bs, 1H), 4.20(m, 3H), 4.05 (s, 3H), 3.22 (m, 1H), 3.20 (d, 1H), 3.18 (s, 1H), 2.80(m, 1H), 2.78 (s, 3H), 2.45 (m, 1H), 2.15 (m, 1H), 1.62 (m, 1H), 1.50(m, 8H) 1.38 (d, 6H), 1.05 (s, 9H). ³¹P (121.4 MHz, CD₃OD):

36.642 LC/MS=925 (M⁺+1)

Example 61 Preparation of Compound 61

^(1H) NMR (300 MHz, CD₃OD): d 8.28 (d, J=9.3 Hz, 1H), 8.18 (s, 1H), 7.76(s, 2H), 7.37 (m, 3H), 7.20 (m, 1H), 5.99 (m, 1H), 5.81 (bs, 1H), 5.29(d, J=17.1 Hz, 1H), 5.11 (d, J=10.5 Hz, 1H), 4.72 (m, 2H), 4.45 (bs,1H), 4.18 (m, 2H), 4.05 (m, 4H), 3.79 (m, 2H), 2.81 (m, 1H), 2.59 (m,1H), 2.21 (m, 1H), 1.68-1.50 (m, 10H) 1.38 (d, J=6.3, 6H), 1.04 (s, 9H).³¹P (121.4 MHz, CD₃OD):

41.451

LC/MS=941 (M⁺+1)

Example 62 Preparation of Compound 62

Intermediate IV (2.06 g, 6.7 mmol) was dissolved in CH₂Cl₂ (60 mL) andcooled to 0° C. Diisopropylethylamine (2.48 mL, 14.3 mmol) was added andstirred for 15 minutes. Chlorotrimethylsilane (1.92 mL, 14.3 mmol) wasadded dropwise. The reaction mixture was warmed to r.t. and stirred for1.5 h. 2-Fluoro-6-trifluoromethylbenzyl chloride (8.61 g, 33.5 mmol) wasadded and the reaction was heated to 45° C. overnight. The reactionmixture was cooled to rt, diluted with CH₂Cl₂, washed with aqueousNH₄Cl, dried with Na₂SO₄, and concentrated. The crude product waspurified by combi-flash to give 1.14 g of phosphinate in 35% yield

To a solution of phosphinate obtained above (550 mg, 1.13 mmol) in CH₃CN(2 mL) at 0° C. was added iodotrimethylsilane (1.13 g, 5.66 mmol). Thereaction mixture was warmed to r.t. and stirred for 30 minutes and thencooled to 0° C. 2,6-Lutidine (0.7 mL) and MeOH (1 mL) were added andstirred for 10 min. The solvent was concentrated and the residue wasco-evaporated with toluene (5 mL), and dried under vacuum for 20 minutesto give crude amine, which was coupled to acid VII to give example 62(339 mg). ¹H NMR (300 MHz, CD₃OD):

8.28 (d, J=9.3 Hz, 1H), 8.18 (s, 1H), 7.76 (m, 2H), 7.54-7.³¹ (m, 4H),5.99 (m, 1H), 5.82 (s, 1H), 5.29 (d, J=17.1 Hz, 1H), 5.13 (m, 1H), 4.71(m, 2H), 4.43 (s, 1H), 4.22-4.05 (m, 2H), 3.78-3.49 (m, 2H), 3.³¹ (m,2H), 2.82 (m, 1H), 2.57 (m, 1H), 2.20 (m, 1H), 1.68-1.48 (m, 8H), 1.34(m, 6H), 1.01 (s, 9H). ³¹P (121.4 MHz, CDCl₃):

40.019

LC/MS=959.37 (M⁺+1), 981.25 (M⁺+Na)

Example 63 Preparation of Compound 63

^(1H) NMR (300 MHz, CD₃OD): d 8.28 (d, J=9.6 Hz, 1H), 8.18 (s, 1H), 7.76(s, 2H), 7.33 (m, 1H), 7.18 (m, 1H), 6.77 (m, 1H), 6.68 (m, 1H), 5.94(m, 1H), 5.81 (bs, 1H), 5.27 (d, J=16.8 Hz, 1H), 5.08 (d, J=9.0 Hz, 1H),4.68 (m, 2H), 4.46 (bs, 1H), 4.17 (m, 2H), 4.05 (m, 4H), 3.81 (m, 3H),3.37 (m, 2H), 2.79 (m, 1H), 2.57 (m, 1H), 2.16 (m, 1H), 1.61-1.50 (m,10H) 1.38 (d, J=6.3 Hz, 6H), 1.04 (s, 9H). ³¹P (121.4 MHz, CD₃OD):

42.834

LC/MS=921 (M⁺+1)

Example 64 Preparation of Compound 64

Intermediate IV (948.8 mg, 3.07 mmol) was dissolved in THF (9.5 mL) andcooled to −40° C. 1 M THF solution of NaN(TMS)₂ (4 mL, 4 mmol) was addeddropwise and the reaction mixture was stirred at −40° C. for 40 minutes.2,6-Dimethylbenzyl chloride (623.2 mg, 4.03 mmol) in THF (2 mL) wasadded and the cold bath was removed. The reaction mixture was stirred atrt for 20 h. The reaction mixture was quenched with 1N HCl (50 mL) andextracted with EtOAc (2×50 mL). The organic layers were washed withbrine, dried with Na₂SO₄, filtered, and concentrated. The crude productwas purified by combi-flash to give 679.9 mg of phosphinate in 52%yield.

A solution of phosphinate (400.1 mg, 0.94 mmol) in CH₃CN (4 mL) wasstirred at 0° C. as iodotrimethylsilane (0.67 mL, 4.71 mmol) was added.The reaction mixture was warmed to r.t. and stirred for 1.5 h. Thereaction mixture was cooled to 0° C. and TEA (10.76 mL) and MeOH (4 mL)were added. The solution was stirred at r.t. for 0.5 h and concentrated.The residue was triturated with toluene (8 mL) and concentrated. Thecrude product was dried and used for next step reaction.

The acid VII (408.5 mg, 0.63 mmol) and amine obtained above weredissolved in DMF (5 mL) and cooled to 0° C. HATU (891.2 mg, 2.34 mmol)and NMM (0.52 mL, 4.73 mmol) were added and the mixture was warmed tor.t. and stirred for 3 h. The crude product was purified by HPLC to give266 mg of 64. ^(1H) NMR (300 MHz, CD₃OD): d 8.26 (d, 1H, J=9.0 Hz), 8.20(s, 1H), 7.74 (br, 2H), 7.30 (dd, 1H, J=9.0 and 2.0 Hz), 6.95 (s, 3H),6.01 (dt, 1H, J=17.1 and 9.8 Hz), 5.80 (br, 1H), 5.29 (dd, 1H, J=17.1and 1.9 Hz), 5.13 (dd, 1H, J=9.8 and 1.9 Hz), 4.62-4.77 (m, 2H), 4.46(br, 1H), 4.05-4.22 (m, 3H), 4.04 (s, 3H), 3.47 (t, 1H, J=15.3 Hz), 3.35(t, 1H, J=15.3 Hz), 2.81 (dd, 1H, J=13.5 and 7.2 Hz), 2.45-2.57 (m, 1H),2.39 (s, 6H), 2.12-2.26 (br m, 1H), 1.39-1.70 (m, 10H), 1.34 (d, 6H,J=6.6 Hz), 1.03 (s, 9H). ³¹P (121.4 MHz, CD₃OD): d 42.678.

LC/MS=901 (M⁺+1)

Example 65 Preparation of Compound 65

Compound IV (3.9 g, 12.6 mmol) was dissolved in CH₂Cl₂ (60 mL) andcooled to 0° C. and diisopropylethylamine (4.5 mL, 26.4 mmol) was added.Chlorotrimethylsilane (3.3 mL, 26.4 mmol) was added dropwise. Thereaction mixture was warmed to r.t. and stirred for 1 h.2,6-difluoro-3-chlorobenzyl bromide (4.5 g, 18.8 mmol) was added and thereaction was heated to 42° C. overnight. The reaction mixture was cooledto rt, diluted with CH₂Cl₂, washed with aqueous NH₄Cl, dried withNa₂SO₄, and concentrated. The crude product was purified by combi-flashto give 3.7 g of phosphinate in 61% yield. The phosphinate was treatedwith TMSI and coupled with VII to give compound 65.

^(1H) NMR (300 MHz, CD₃OD): d 8.23 (d, J=9.5 Hz, 1H), 8.20 (s, 1H), 7.79(s, 2H), 7.36 (m, 2H), 6.97 (m, 1H), 5.95 (m, 1H), 5.78 (bs, 1H), 5.32(d, J=9.6 Hz, 1H), 5.18 (d, J=9.0 Hz, 1H), 4.63 (m, 2H), 4.45 (bs, 1H),4.20 (m, 3H), 4.05 (s, 3H), 3.34 (m, 2H), 2.80 (m, 1H), 2.45 (m, 1H),2.15 (m, 1H), 1.68-1.50 (m, 8H) 1.38 (d, 6H), 1.05 (s, 9H). ³¹P (121.4MHz, CD₃OD):

40.079

LC/MS=943 (M⁺+1)

Example 66 Preparation of Compound 66

^(1H) NMR (300 MHz, CD₃OD): d 8.23 (d, J=9.5 Hz, 1H), 8.20 (s, 1H), 7.79(s, 2H), 7.36 (m, 1H), 7.06 (m, 2H), 5.95 (m, 1H), 5.78 (bs, 1H), 5.32(d, J=9.6 Hz, 1H), 5.18 (d, J=9.0 Hz, 1H), 4.63 (m, 2H), 4.45 (bs, 1H),4.20 (m, 3H), 4.05 (s, 3H), 3.34 (m, 2H), 2.80 (m, 1H), 2.45 (m, 1H),2.15 (m, 1H), 1.68-1.50 (m, 8H) 1.38 (d, 6H), 1.05 (s, 9H). ³¹P (121.4MHz, CD₃OD):

40.879

LC/MS=943 (M⁺+1)

Example 67 Preparation of Compound 67

¹H NMR (300 MHz, CD₃OD): d 8.28 (d, J=9.3 Hz, 1H), 8.17 (s, 1H), 7.75(m, 2H), 7.68 (m, 1H), 7.32 (d, J=9.3 Hz, 1H), 7.08 (dd, J=9.6, 9.6 Hz,1H), 5.94-5.82 (m, 2H), 5.79 (s, 1H), 5.26 (d, J=17.1 Hz, 1H), 5.05 (d,J=10.2 Hz, 1H), 4.65 (m, 2H), 4.15 (m, 3H), 4.05 (s, 3H), 3.³¹ (m, 2H),2.80 (m, 1H), 2.51 (m, 1H), 2.11 (m, 1H), 1.63-1.49 (m, 8H), 1.34 (m,6H), 1.00 (s, 9H). ³¹P (121.4 MHz, CDCl₃):

40.908 LC/MS=943.27 (M⁺+1), 965.03 (M⁺+Na)

Example 68 Preparation of Compound 68

¹H NMR (300 MHz, CD₃OD) d 8.28 (d, J=9.3 Hz, 1H), 8.16 (s, 1H), 7.77 (s,1H), 7.72 (d, J=2.1 Hz, 1H), 7.33 (d, J=12 Hz, 1H), 7.14 (m, 1H), 6.91(m, 1H), 5.98 (dt, J=10.2, 17.1 Hz, 1H), 5.79 (s, 1H), 5.³¹ (d, J=16.8Hz, 1H), 5.13 (d, J=11.7 Hz, 1H), 4.71 (t, J=9 Hz), 4.63 (d, J=11.1 Hz,1H), 4.5 (s, 1H), 4.1-4.2 (brm, 3H), 4.05 (s, 3H), 3.44 (dd, J=5.1, 15.6Hz, 2H), 2.77 (m, 1H), 2.59 (m, 1H), 2.19 (m, 1H), 1.44-1.7 (m, 10H),1.34 (d, J=6.3 Hz, 6H), 1.04 (s, 9H). ³¹P NMR (121.4 MHz, CD₃OD) d 37.8

LC/MS=927.3 (M⁺+1)

Example 69 Preparation of Compound 69

Intermediate IV (15.5 g, 50.3 mmol) was dissolved in CH₂Cl₂ (300 mL) andcooled to 0° C. and diisopropylethylamine (22 mL, 126 mmol) was added.Chlorotrimethylsilane (17 mL, 126 mmol) was added dropwise. The reactionmixture was warmed to r.t. and stirred for 1 h. 2,3,6-Trifluorobenzylbromide (37 g, 165 mmol) was added and the reaction was stirred at r.r.overnight. Aqueous NH₄Cl (200 mL) was added and stirred for 30 minutes.The two layers were separated and aqueous layer was extracted withCH₂Cl₂. The combined organic layer was dried with Na₂SO₄ andconcentrated. The crude product was purified by combi-flash to give 7.3g of phosphinate.

To a solution of phosphinate (7.2 g, 15.8 mmol) in TFA (45 mL) at r.t.was added DMS (10 mL) and stirred overnight. The mixture wasconcentrated and co-evaporated with toluene. The residue was dissolvedin 1/1 iPrOH/heptane and washed with 6 N HCl (3×). The combined aqueouslayers were brought to pH=10 with NaOH in a cold bath. The aqueous layerwas extracted with EtOAc (3×100 mL). The organic layers were washed withbrine, dried with Na₂SO₄, and concentrated to give 3.8 g of amine whichwas coupled and deprotected to give compound 69 in 75% yield.

¹H NMR (300 MHz, CD₃OD):

8.27 (d, J=9.3 Hz, 1H), 8.18 (s, 1H), 7.76 (d, J=3.0 Hz, 2H), 7.43 (m,1H), 7.28 (d, J=2.4 Hz, 1H), 7.03 (dd, J=8.7, 8.7 Hz, 1H), 5.91 (m, 1H),5.81 (s, 1H), 5.27 (d, J=17.1 Hz, 1H), 5.08 (d, J=11.4 Hz, 1H), 4.68 (m,2H), 4.46 (s, 1H), 4.16 (m, 3H), 3.36 (m, 2H), 2.80 (m, 1H), 2.55 (m,1H), 2.13 (m, 1H), 1.62-1.46 (m, 8H), 1.34 (d, J=6.3 Hz, 6H), 1.02 (s,9H). ³¹P (121.4 MHz, CDCl₃): d 41.986. LC/MS=943.27 (M⁺+1), 965.03(M⁺+Na)

Example 70 Preparation of Compound 70

Intermediate IV (380 mg, 0.78 mmol) was dissolved in CH₂Cl₂ (15 mL) andcooled to 0° C. Diisopropylethylamine (0.4 mL, 2.3 mmol) was added andstirred for 25 minutes. Chlorotrimethylsilane (0.32 mL, 2.3 mmol) wasadded dropwise. The reaction mixture was warmed to r.t. and stirred for1 h. 1-Bromomethyl-5-chloro-2,4-difluoro-benzene (940 mg, 3.9 mmol) wasadded and the reaction was heated to 45° C. for 18 h. The reactionmixture was cooled to rt, diluted with CH₂Cl₂, washed with aqueousNH₄Cl, dried with Na₂SO₄, and concentrated. The crude product waspurified by combi-flash to give 190 mg of phosphinate in 52% yield.

To a solution of phosphinate obtained above (190 mg, 0.41 mmol) in CH₃CN(1 mL) at 0° C. was added iodotrimethylsilane (0.3 mL, 2 mmol). Thereaction mixture was warmed to r.t. and stirred for 30 minutes and thencooled to 0° C. 2,6-Lutidine (0.23 mL) and MeOH (1 mL) were added andstirred for 10 minutes. The solvent was concentrated and the residue wasco-evaporated with toluene (5 mL), and dried under vacuum for 20 minutesto give crude amine. Coupling with acid VII (130 mg, 0.2 mmol) providedcompound 70. ^(1H) NMR (300 MHz, CD₃OD): d 8.23 (d, J=9.5 Hz, 1H), 8.20(s, 1H), 7.79 (s, 2H), 7.36 (m, 1H), 7.14 (m, 2H), 5.95 (m, 1H), 5.78(bs, 1H), 5.32 (d, J=9.6 Hz, 1H), 5.18 (d, J=9.0 Hz, 1H), 4.63 (m, 2H),4.45 (bs, 1H), 4.20 (m, 3H), 4.05 (s, 3H), 3.34 (m, 2H), 2.80 (m, 1H),2.45 (m, 1H), 2.15 (m, 1H), 1.68-1.50 (m, 8H) 1.38 (d, 6H), 1.05 (s,9H). ³¹P (121.4 MHz, CD₃OD):

0.578. LC/MS=943 (M⁺+1)

Example 71 Preparation of Compound 71

^(1H) NMR (300 MHz, CD₃OD): d 8.28 (d, J=9.6 Hz, 1H), 8.18 (s, 1H), 7.76(s, 2H), 7.44 (m, 1H), 7.34 (m, 1H), 7.08 (t, d=9.0 Hz, 1H), 5.95 (m,1H), 5.80 (bs, 1H), 5.29 (d, J=17.4 Hz, 1H), 5.13 (d, J=9.0 Hz, 1H),4.69 (m, 2H), 4.47 (bs, 1H), 4.18 (m, 2H), 4.06 (m, 4H), 3.58 (m, 2H),2.79 (m, 1H), 2.57 (m, 1H), 2.19 (m, 1H), 1.68-1.50 (m, 10H) 1.38 (d,J=6.3 Hz, 6H), 1.04 (s, 9H). ³¹P (121.4 MHz, CD₃OD):

40.778. LC/MS=959 (M⁺+1)

Example 72 Preparation of Compound 72

The phosphonous acid IV (1.62 g, 5.27 mmol) was suspended in 5 mL of THFand cooled to −40° C. 1N NaN(TMS)₂ (6.32 mL, 6.³¹ mmol) was addeddropwise over 15 minutes followed by 1-(bromomethyl)-2-nitrobenzene(1.36 g, 6.32 mmol) in 1 mL of THF. The solution stirred from −40° C. tort overnight. The reaction was diluted with EtOAc and quenched with 20mL of 1N HCl. The organic layer was washed with brine, dried over MgSO₄,filtered and concentrated. The crude material was purified using aCombiFlash Chromatography System using a gradient of 30% EtOAc/Hex to100% EtOAc to obtain(1-Benzyloxycarbonylamino-2-vinyl-cyclopropyl)-(2-nitro-benzyl)-phosphinicacid ethyl ester (196 mg, 8%) as a brown oil.

The phosphinate (196 mg, 0.44 mmol) was suspended in 1 mL of CH₃CN andcooled to 0° C. Iodotrimethylsilyl (TMSI) (155 μl, 1.08 mmol) was addedand the solution was warmed to rt. After 45 minutes, the solution wascooled again to 0° C. and triethylamine (1 mL, 7.33 mmol) and 2 mL ofMeOH. The solution was warmed to rt and stirred for an additional 20minutes. The solution was concentrated, azeotroped 2× with toluene andput on high vacuum for 30 minutes to provide(1-Amino-2-vinyl-cyclopropyl)-(2-nitro-benzyl)-phosphinic acid. The acid(124 mg, 0.44 mmol) was coupled with intermediate IV (191 mg, 0.29mmol), HATU (276 mg, 0.73 mmol), and NMM (160 μl, 1.45 mmol) to givecompound 72 (140 mg, 53%) as a yellow solid. ^(1H) NMR (300 MHz, CD₃OD):d 8.30 (d, J=9.5 Hz, 1H), 8.20 (s, 1H), 7.79 (s, 2H), 7.35 (d, J=9.3 Hz,1H), 7.33 (m, 1H), 6.94 (m, 2H), 5.95 (m, 1H), 5.80 (s, 1H), 5.25 (d,J=9.6 Hz, 2H), 5.17 (d, J=9.0 Hz, 2H), 4.75 (m, 2H), 4.45 (bs, 1H), 4.20(s, 2H), 4.05 (s, 3H), 3.40 (m, 2H), 2.80 (m, 1H), 2.52 (m, 1H), 2.15(m, 1H), 1.62 (m, 6H), 1.38 (d, 6H), 1.05 (s, 9H). ³¹P (121.4 MHz,CD₃OD):

40.898

Example 73 Preparation of Compound 73

Compound 72 (80 mg, 0.08 mmol) was suspended in EtOH and SnCl₂2H₂O (98mg, 0.44 mmol) was added. The solution was heated to reflux. After 3hours, the starting material was consumed. The solution was filtered andconcentrated. The mixture was purified via Gilson HPLC to obtain 73 (20mg, 53%) as a yellow solid. ^(1H) NMR (300 MHz, CD₃OD): d 8.30 (d, J=9.5Hz, 1H), 8.20 (s, 1H), 7.79 (s, 2H), 7.35 (d, J=9.3 Hz, 1H), 7.33 (m,1H), 6.94 (m, 2H), 5.95 (m, 1H), 5.80 (s, 1H), 5.25 (d, J=9.6 Hz, 2H),5.17 (d, J=9.0 Hz, 2H), 4.75 (m, 2H), 4.45 (bs, 1H), 4.20 (s, 2H), 4.05(s, 3H), 3.40 (m, 2H), 2.80 (m, 1H), 2.52 (m, 1H), 2.15 (m, 1H), 1.62(min, 6H), 1.38 (d, 6H), 1.05 (s, 9H). ³¹P (121.4 MHz, CD₃OD):

40.898

Example 74 Preparation of Compound 74

A solution of compound IV (96 mg, 0.³¹ mmol) in CH₂Cl₂ (2.82 mL) wasadded DIEA (0.114 mL, 0.652 mmol) and TMSCl (0.083 mL, 0.652 mmol) at 0°C. The reaction was allowed to warm up to rt and stirred for 1 hour. Tothe mixture was added a solution of 2-(bromomethyl)pyridine (173 mg,0.683 mmol) in DIEA (0.054 mL, 0.³¹ mmol). This reaction was stirred atrt for 2 days when complete consumption of the starting materials wasobserved by LCMS. The reaction was worked up addition of CH₂Cl₂ andsaturated aqueous NH₄Cl. The organic layer was dried in vacuo andpurified using silica gel chromatography to give 91 mg of the product asa clear oil. EI MS (m/z) 401.0 [M+H].

A solution of benzyl(1S,2S)-1-((S)-ethoxy(pyridin-2-ylmethyl)phosphoryl)-2-vinylcyclopropylcarbamate(96 mg, 0.239 mmol) in 2.39 mL of aqueous 6N HCl was heated at 70° C.for 7 hours and stirred at rt for 12 hours. The reaction mixture wasworked up by removal of all volatiles and was carried on without anyfurther purification. EI MS (m/z) 267.3 [MH⁺].

A solution of(S)-ethyl((1S,2S)-1-amino-2-vinylcyclopropyl)(pyridin-2-ylmethyl)phosphinate(64 mg, 0.24 mmol), carboxylic acid VII (157 mg, 0.0.24 mmol) in a 1:1solution of DMF-CH₂Cl₂ (1.2 mL) was stirred with HATU (137 mg, 0.36mmol) and DIEA (0.168 mL, 0.962 mmol) for 2.5 hours when the reactionwas complete. The product was purified by silica gel chromatography(EtOAc-EtOAc/MeOH) to provide 82 mg of the desired product. EI MS (m/z)901.4 [MH⁺].

A solution ofcyclopentyl(S)-1-((2S,4R)-2-(((1S,2S)-1-((S)-ethoxy(pyridin-2-ylmethyl)phosphoryl)-2-vinylcyclopropyl)-carbamoyl)-4-(2-(2-(isopropylamino)thiazol-4-yl)-7-methoxyquinolin-4-yloxy)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-ylcarbamate(68 mg, 0.074 mmol) and TMSI (0.053 mL, 0.371 mmol) was stirred in dryacetonitrile (0.74 mL) for 1 hour when the reaction was complete asjudged by LCMS. The reaction was quenched using TEA (0.104 mL, 0.742mmol) followed by addition of MeOH (10 mL). The reaction mixture wasdried under reduced pressure and the residue was purified by RP HPLC(ACN, 0.05% TFA-H₂O, 0.05% TFA) to provide 42 mg of the desired product.¹H NMR (300 MHz, CD₃OD) d 8.60 (d, 1H, J=5 Hz), 8.44 (t, 1H, J=9 Hz),8.³¹ (d, 1H, J=9 Hz), 8.17 (s, 1H), 7.98 (d, 1H, J=9 Hz), 7.86 (t, 1H,J=6 Hz), 7.76 (s, 2H), 7.33 (d, 1H, J=11 Hz), 5.83 (br s, 1H), 5.55 (dt,1H, J=9, 17 Hz), 4.98 (d, 1H, J=17 Hz), 4.78-4.65 (m, 2H), 4.66-4.51 (m,2H), 4.21-4.07 (m, 3H), 4.05 (s, 3H), 3.54 (d, 1H, J=8 Hz), 3.48 (d, 1H,J=6 Hz), 2.87-2.82 (m, 1H), 2.61-2.45 (m, 1H), 2.08-1.94 (m, 1H),1.70-1.44 (m, 8H), 1.5-1.35 (m, 2H), 1.34 (d, 6H, J=7 Hz), 1.08 (s, 9H);³¹P (121.4 MHz, CD₃OD)

23.5; EI MS (m/z) 873.7 [MH⁺].

Example 75 Preparation of Compound 75

A solution of compound IV (161 mg, 0.521 mmol) in CH₂Cl₂ (4.7 mL) wasadded DIEA (0.190 mL, 1.09 mmol) and TMSCl (0.139 mL, 1.09 mmol) at 0°C. The reaction was allowed to warm up to rt and stirred for 1 hour. Tothe mixture was added a solution of 3-(bromomethyl)pyridine (290 mg,1.15 mmol) in DIEA (0.091 mL, 0.521 mmol). This reaction was stirred atrt for 3 days when complete consumption of the starting materials wasobserved by LCMS. The reaction was worked up addition of CH₂Cl₂ andsaturated aqueous NH₄Cl. The organic layer was dried in vacuo andpurified using silica gel chromatography to give 91 mg of the product asa clear oil. EI MS (m/z) 401.0 [M+H].

A solution of benzyl(1S,2S)-1-((S)-ethoxy(pyridin-3-ylmethyl)phosphoryl)-2-vinylcyclopropylcarbamate(41 mg, 0.102 mmol) in acetonitrile (1.02 mL) was treated with TMSI(0.073 mL, 0.512 mmol) for 2 hours at rt when the reaction was complete.The reaction was quenched by addition of TEA (0.142 mL, 1.02 mmol) andMeOH (10 mL) and the residue was dried and used as is.

A solution(S)-((1S,2S)-1-amino-2-vinylcyclopropyl)(pyridin-3-ylmethyl)phosphinicacid (24 mg, 0.10 mmol), carboxylic acid VII (66 mg, 0.100 mmol) in DMF(1.0 mL) was stirred with HATU (57 mg, 0.15 mmol) and DIEA (0.070 mL,0.403 mmol) for 1 hour when the reaction was complete. The product waspurified by RP HPLC (ACN, 0.05% TFA-H₂O, 0.05% TFA) to provide 28 mg ofthe desired product. ¹H NMR (300 MHz, CD₃OD) d 8.89 (s, 1H), 8.61 (d,1H, J=5 Hz), 8.52 (d, 1H, J=8 Hz), 8.30 (d, 1H, J=9 Hz), 8.17 (s, 1H),7.90 (t, 1H, J=6 Hz), 7.76 (s, 2H), 7.32 (d, 1H, J=10 Hz), 5.80 (br s,1H), 5.77-5.65 (m, 1H), 5.07 (d, 1H, J=17 Hz), 4.79 (d, 1H, J=11 Hz),4.71-4.63 (m, 2H), 4.49 (br s, 1H), 4.23-4.09 (m, 3H), 4.05 (s, 3H),3.46-3.23 (m, 2H), 2.90-2.78 (m, 1H), 2.57-2.46 (m, 1H), 2.07-1.93 (m,1H), 1.70-1.43 (m, 8H), 1.43-1.30 (m, 2H), 1.34 (d, 6H, J=6 Hz), 1.03(s, 9H); ³¹P (121.4 MHz, CD₃OD)

31.7; EI MS (m/z) 874.0 [MH⁺].

Example 76 Preparation of Compound 76

A solution of compound IV (228 mg, 0.737 mmol) in CH₂Cl₂ (6.7 mL) wasadded DIEA (0.270 mL, 1.55 mmol) and TMSCl (0.196 mL, 1.55 mmol) at 0°C. The reaction was allowed to warm to rt and was stirred for 1 hour. Tothe mixture was added 2-bromomethyl-3-hydroxypyridine hydrochloride (436mg, 1.62 mmol) in DIEA (0.128 mL, 0.737 mmol). This reaction was stirredat room temperature for 1 day, and CH₂Cl₂ and saturated aqueous NH₄Clwere added. The organic layer was dried in vacuo and purified usingsilica gel chromatography to give 205 mg (67%) of the product as a clearoil. EI MS (m/z) 439.0 [M+H].

A solution of benzyl(1S,2S)-1-((S)-ethoxy((3-hydroxypyridin-2-yl)methyl)phosphoryl)-2-vinylcyclopropyl-carbamate(205 mg, 0.492 mmol) in acetonitrile (4.92 mL) was treated with TMSI(0.350 mL, 2.46 mmol) for 2 hours at rt when the reaction was complete.The reaction was quenched by addition of TEA (0.685 mL, 4.92 mmol) andMeOH (10 mL) and the residue was dried and used as is.

A solution of(S)-((1S,2S)-1-amino-2-vinylcyclopropyl)((3-hydroxypyridin-2-yl)methyl)phosphinicacid (214 mg, 0.328 mmol), carboxylic acid VII (125 mg, 0.493 mmol) inDMF (1.5 mL) was stirred with HATU (188 mg, 0.493 mmol) and DIEA (0.228mL, 1.30 mmol) for 1 hour when the reaction was complete. The productwas purified by RP HPLC (ACN, 0.05% TFA-H₂O, 0.05% TFA) to provide 54 mgof the desired product. ¹H NMR (300 MHz, CD₃OD) d 8.29 (d, 1H, J=9 Hz),8.17 (s, 1H), 8.11 (d, 1H, J=8 Hz), 7.82 (d, 1H, J=8 Hz), 7.76 (s, 2H),7.69-7.61 (m, 1H), 7.³¹ (d, 1H, J=9 Hz), 5.28 (br s, 1H), 5.71 (dt, 1H,J=10, 17 Hz), 5.04 (d, 1H, J=17 Hz), 4.79-4.63 (m, 2H), 4.50 (br s, 1H),4.25-4.05 (m, 3H), 4.05 (s, 3H), 3.68 (app t, 1H, J=15 Hz), 3.41 (t, 1H,J=16 Hz), 2.95-2.84 (m, 1H), 2.60-2.48 (m, 1H), 2.08-1.97 (m, 1H),1.70-1.45 (m, 8H), 1.45-1.35 (m, 2H), 1.34 (d, 6H, J=7 Hz), 1.03 (s,9H); ³¹P (121.4 MHz, CD₃OD)

26.7; EI MS (m/z) 889.7 [MH⁺].

Example 77 Preparation of Compound 77

A solution of 3-chloro-6-methylpyridine (220 mg, 1.72 mmol) incarbontetrachloride (4 mL) was heated with NBS (284 mg, 1.60 mmol) andbenzoyl peroxide (100 mg) for 3 days. The reaction was worked up byremoval of the solvent and resuspension of the production in CH₂Cl₂. Theorganic layer was washed with aqueous 2N NaOH (2×50 mL) and dried invacuo to give 170 mg of the product as a clear oil; EI MS (m/z) 208.0,210.0 [M+H].

A solution of compound IV (102 mg, 0.330 mmol) in CH₂Cl₂ (3.0 mL) wasadded DIEA (0.121 mL, 0.692 mmol) and TMSCl (0.088 mL, 0.692 mmol) at 0°C. The reaction was allowed to warm up to rt and stirred for 1 hour. Tothe mixture was added 2-(bromomethyl)-6-chloropyridine (102 mg, 0.330mmol) in DIEA (0.121 mL, 0.692 mmol). This reaction was stirred at rtover night when it was worked up by addition of CH₂Cl₂ and saturatedaqueous NH₄Cl. The organic layer was dried in vacuo and purified usingsilica gel chromatography to give 140 mg (97%) of the product as a clearoil. EI MS (m/z) 457.0 [M+Na].

A solution benzyl(1S,2S)-1-((S)-((6-chloropyridin-2-yl)methyl)(ethoxy)phosphoryl)-2-vinylcyclopropylcarbamate(118 mg, 0.271 mmol) in acetonitrile (2.71 mL) was treated with TMSI(0.193 mL, 1/35 mmol) for 1.5 hours at rt when the reaction wascomplete. The reaction was quenched by addition of TEA (0.377 mL, 2.71mmol) and MeOH (10 mL) and the residue was dried and used as is; EI MS(m/z) 273.1 [MH⁺].

A solution of(S)-((1S,2S)-1-amino-2-vinylcyclopropyl)((6-chloropyridin-2-yl)methyl)phosphinicacid (74 mg, 0.271 mmol), carboxylic acid VII (177 mg, 0.271 mmol) inDMF (1.3 mL) was stirred with HATU (155 mg, 0.407 mmol) and DIEA (0.189mL, 1.09 mmol) for 1 hour when the reaction was complete. The productwas purified by RP HPLC (ACN, 0.05% TFA-H₂O, 0.05% TFA) to provide 37 mgof the desired product. ¹H NMR (300 MHz, CD₃OD) d 8.29 (d, 1H, J=9 Hz),8.17 (s, 1H), 7.77-7.65 (m, 3H), 7.43 (dd, 1H, J=2, 8 Hz), 7.35-7.27 (m,2H), 5.92-5.75 (m, 2H), 5.23 (d, 1H, J=17 Hz), 5.01 (d, 1H, J=12 Hz),4.75-4.61 (m, 2H), 4.50 (br s, 1H), 4.20-4.08 (m, 3H), 4.05 (s, 3H),3.53 (dd, 2H, J=3, 17 Hz), 2.84-2.74 (m, 1H), 2.65-2.53 (m, 1H),2.16-2.04 (m, 1H), 1.70-1.42 (m, 10H), 1.34 (d, 6H, J=6 Hz), 1.03 (s,9H); ³¹P (121.4 MHz, CD₃OD)

40.7; EI MS (m/z) 907.4 [MH⁺].

Example 78 Preparation of Compound 78

Examples 78 through 81 were prepared in a manner similar to that used toprepare example 74.

The product (Example 78) was afforded as a yellow solid, (48 mg, 15%).^(1H) NMR (300 MHz, CD₃OD): d 8.73 (s, 1H), 8.53 (s, 1H), 8.44 (s, 1H)8.28 (d, J=9.2 Hz, 1H) 8.195 (s, 1H) 7.76 (s, 2H) 7.³¹ (d, J=8.8 Hz, 1H)5.84 (m, 2H), 5.20 (m, 1H), 4.99 (m, 1H), 4.71 (m, 2H), 4.48 (bs, 1H),4.15 (m, 3H) 4.04 (s, 3H), 3.60 (m, 2H), 2.75 (m, 1H), 2.54 (m, 1H),2.02 (m, 1H), 1.54 (m, 8H) 1.34 (m, 8H), 1.01 (s, 9H). ³¹P (121.4 MHz,CD₃OD):

38.710. LC (6 minute run, r.t.=3.50 min) MS (875.5, M+1)

Example 79 Preparation of Compound 79

The product, (Example 79) was afforded as a yellow solid (7 mg, 5%).

^(1H) NMR (300 MHz, CD₃OD): d 9.00 (s, 1H) 8.82 (s, 2H) 8.29 (d, J=8.8Hz, 1H) 8.18 (s, 1H) 7.75 (s, 2H), 7.32, (d, J=8.1 Hz, 1H) 5.80 (m, 2H),5.18 (m, 1H), 4.95 (m, 1H), 4.66 (m, 2H), 4.47 (bs, 1H), 4.18 (m, 3H),4.05 (s, 3H) 2.77 (m, 1H), 2.49 (m, 1H), 2.06 (m, 1H), 1.50 (m, 8H),1.34 (m, 8H), 1.01 (s, 9H). Not enough material for ³¹P NMR. LC (6minute run, r.t.=3.42 min) MS (875.5, M+1)

Example 80 Preparation of Compound 80

The product, (Example 80) was afforded as a yellow solid (11 mg, 15

^(1H) NMR (300 MHz, CD₃OD): d 8.74 (d, J=4.9 Hz, 2H) 8.29 (d, J=9.4 Hz,1H) 8.17 (s, 1H) 7.76 (m, 2H), 7.35 (m, 2H) 5.86 (m, 2H), 5.22 (m, 1H),5.00 (m, 1H), 4.70 (m, 2H), 4.49 (bs, 1H), 4.17 (m, 3H), 4.05 (s, 3H)3.70 (m, 2H) 2.78 (m, 1H), 2.59 (m, 1H), 2.12 (m, 1H), 1.59 (m, 8H),1.34 (m, 8H), 1.02 (s, 9H).

³¹P (121.4 MHz, CD₃OD):

37.909. LC (6 minute run, r.t.=3.21 min) MS (875.6, M+1)

Example 81 Preparation of Compound 81

The product (Example 81) was afforded as a yellow solid (85 mg, 51%).

^(1H) NMR (300 MHz, CD₃OD): d 9.02 (s, 1H) 8.65 (d, J=5.2 Hz, 1H) 8.28(d, J=9.5 Hz, 1H) 8.18 (s, 1H) 7.75 (m, 2H), 7.66, (d, J=5.0 Hz, 1H)7.30 (m, 1H) 5.86 (m, 2H), 5.20 (m, 1H), 5.00 (m, 1H), 4.68 (m, 2H),4.47 (bs, 1H), 4.17 (m, 3H), 4.05 (s, 3H) 3.57 (m, 2H) 2.78 (m, 1H),2.56 (m, 1H), 2.08 (m, 1H), 1.60 (m, 8H), 1.34 (m, 8H), 1.02 (s, 9H).³¹P (121.4 MHz, CD₃OD): d 36.81. LC (6 minute run, r.t.=3.21 min) MS(875.5, M+1)

Example 82 Preparation of Compound 82

¹H NMR (300 MHz, CD₃OD) 8.27 (d, J=9.0 Hz, 1H), 8.20 (s, 1H), 7.76 (s,2H), 7.³¹ (b, 1H), 7.221 (b, 1H), 7.00 (b, 1H), 6.93 (m, 1H), 5.95 (m,1H), 5.80 ((b, 1H), 5.24 (d, J=17.4 Hz, 1H), 5.07 (d, J=10.2 Hz, 1H),4.68 (m, 2H), 4.46 (s, 1H), 4.17 (m, 2H), 4.11 (s, 1H), 4.04 (s, 3H),3.49 (d, 15 Hz, 2H), 2.75 (m, 1H), 2.47 (m, 1H), 2.08 (m, 1H), 1.41-1.62(m, 8H), 1.34 (d, J=6.3 Hz, 6H), 1.03 (s, 9H). ³¹P NMR (121.4 MHz,CD₃OD) d 39.122

LC/MS=879 (M⁺+1)

Example 83 Preparation of Compound 83

The furfuryl bromide was formed in situ from the furfuryl alcohol in thefollowing manner. Furfuryl alcohol 3.5 mL (41 mmol) was dissolved in 20mL of dry ether and cooled to 0° C. PBr₃ (1.4 mL, 15.1 mmol) dissolvedin 4 mL of dry ether was then added at 0° C. After addition, thesolution was allowed to warm to rt. After 45 min. at rt, the solutionwas cooled to 0° C. and 12 mL of 50% aqueous KOH solution was added. Theether layer was then decanted into a dry flask and stored at −20° C.over solid KOH.

In a separate flask, 392 mg (1.27 mmol) of IV was dissolved in 5.0 mL ofdry DCM. 465 μl (2.67 mmol) of DIEA and 339 μl (2.67 mmol) of TMSCl wereadded respectively and the reaction then stirred at rt for 5 min. 465 μl(2.67 mmol) of DIEA and 1.7 mL of the ether solution of in situ formedfurfurylbromide mentioned above was then added. The reaction was warmedto 40° C. and allowed to go at 40° C. overnight. The reaction was thendiluted with ethyl acetate and concentrated to remove DCM. The organicphase was then washed with 1× w/1.0 M Citric Acid, 2× w/water, and 1×w/Brine. The organic phase was dried over MgSO₄. Concentration of thefiltrate from vacuum filtration removal of the MgSO₄ yielded an orangeoil from which product 7 was isolated by column chromatography (SiO₂,3:1-Ethylacetate:Hexane) as a clear oil (160 mg, 32% over 2 steps). ¹HNMR (300 MHz, CDCl₃) 7.33 (s, 5H), 6.³¹ (m, 2H), 6.00 (m, 1H), 5.30 (m,2H), 5.04 (m, 4H), 4.10 (m, 2H), 3.35 (m, 2H), 1.96 (m, 2H)), 1.80 (m,1H), 1.60 (m, 1H), 1.303 (m, 3H). ³¹P NMR (121.4 MHz, CDCl₃) d 44.879,41.575. LC/MS=390 (M⁺+1).

A solution of phosphinate obtained above (103 mg, 0.308 mmol) in ACN(7.7 mL) was cooled to 0° C. and TMSI (220 μL, 1.54 mmol) was added in adrop-wise fashion. The reaction was warmed to rt and stirred at rt forone hour. The reaction was cooled back to 0° C. and 2,6-lutidine (360μL, 3.1 mmol) was added in a drop-wise fashion. This was followed by theaddition of Et₃N (1 mL, 7.2 mmol) and MeOH (4 mL). The reaction was thenconcentrated in vacuum and crude was used directly in the next reaction.

Crude residue from step 1, HATU (190 mg, 0.5 mmol), dipeptide VII (130mg, 0.2 mmol) and n-methylmorpholine (110 μl, 1.0 mmol) were dissolvedin 2 mL of DMF and stirred at rt overnight. The crude reaction mixturewas then purified by reverse prep HPLC directly to afford 60 mg of 83(60 mg, 34%)

¹H NMR (300 MHz, CD₃OD) 8.82 (s, 1H), 8.26 (d, J=9.6 Hz, 1H), 8.20 (s,1H), 7.75 (s, 2H), 7.35 (s, 1H), 7.29 (dd, J=2.1, 9.3 Hz, 1H), 6.30 (m,2H), 5.95 (m, 1H), 5.80 ((b, 1H), 5.24 (d, J=11.4 Hz, 1H), 5.07 (d, J=12Hz, 1H), 4.65 (m, 2H), 4.45 (s, 1H), 4.17 (m, 2H), 4.11 (s, 1H), 4.04(s, 3H), 3.35 (m, 2H), 2.80 (m, 1H), 2.50 (m, 1H), 2.10 (m, 1H),1.41-1.78 (m, 8H), 1.34 (d, J=6.3 Hz, 6H), 1.04 (s, 9H). ³¹P NMR (121.4MHz, CD₃OD) d 40.029

LC/MS=863 (M⁺+1)

Example 84 Preparation of Compound 84

Intermediate IV (360 mg, 1.2 mmol) was dissolved in 5.0 mL of dry DCM.DIEA (418 μl, 2.4 mmol) and 343 μl (2.4 mmol) of TMSCl were addedsequentially and the reaction then stirred at rt for 5 min. More DIEA(418 μl, 2.4 mmol) and 343 μl (2.4 mmol) of 5-(trifluoromethyl)furfurylbromide were then added respectively. The reaction was warmed to 40° C.and allowed to stir at 40° C. overnight. The reaction was then dilutedwith ethyl acetate and concentrated to remove DCM. The organic phase wasthen washed with 1× with sat. NH4Cl, 2× with water, and 1× w/brine. Theorganic phase was dried over MgSO₄. Concentration of the filtrate afterfiltration of the MgSO₄ yielded an orange oil from which product wasisolated by column chromatography (SiO₂, neat ethyl acetate) as a clearoil (³¹3 mg, 56%). Deprotection and coupling to dipeptide VII affordedcompound 84.

¹H NMR (300 MHz, CD₃OD) 8.27 (d, J=8.7 Hz, 1H), 8.20 (s, 1H), 7.75 (s,2H), 7.35 (s, 1H), 7.29 (d, J=2.1, 9.3 Hz, 1H), 6.86 (b, 1H), 6.48 (b,1H), 5.90 (b, 1H), 5.79 (b, 1H), 5.25 (d, J=17.4 Hz, 1H), 5.07 (d,J=10.8 Hz, 1H), 4.67 (m, 2H), 4.45 (s, 1H), 4.16 (m, 2H), 4.11 (s, 1H),4.04 (s, 3H), 3.43 (m, 2H), 2.80 (m, 1H), 2.50 (m, 1H), 2.10 (m, 1H),1.62-1.33 (m, 8H), 1.34 (d, J=6.3 Hz, 6H), 1.04 (s, 9H). ³¹P NMR (121.4MHz, CD₃OD) d 36.68

LC/MS=9³¹ (M⁺−1)

Example 85 Preparation of Compound 85

To a solution of 5-methyl-1H-pyrazole (5 g, 61.05 mmol) in CH₃CN (50 mL)at 0° C. was added di-tert-butyl dicarbonate (16 g, 73.26 mmol) and DMAP(740 mg, 6.10 mmol). The reaction mixture was warmed to r.t. and stirredfor 2 h. The reaction mixture was diluted with EtOAc (30 mL) and washedwith 1 N HCl (2×30 mL). The organic layer was washed with saturatedNaHCO₃ (30 mL) and brine (30 mL), dried with Na₂SO₄, and concentrated togive 8.7 g of 5-methyl-pyrazole-1-carboxylic acid tert-butyl ester ascrude product.

To a solution of 5-methyl-pyrazole-1-carboxylic acid tert-butyl ester inCCl₄ (40 mL) was added NBS (3.3 g, 18.5 mmol) and benzoylperoxide (450mg, 1.86 mmol). The reaction mixture was heated to reflux for 4 h andcooled to rt. The insoluble material was filtered off and the solutionwas diluted with EtOAc. The organics were washed with saturated NaHCO₃and H₂O, dried with Na₂SO₄, and concentrated. The crude product waspurified by combi-flash to give 1.67 g of5-bromomethyl-pyrazole-1-carboxylic acid tert-butyl ester in 52% yield.

Intermediate IV (800 mg, 2.56 mmol) was dissolved in CH₂Cl₂ (30 mL) andcooled to 0° C. Diisopropylethylamine (1 mL, 5.36 mmol) was added andstirred for 15 minutes. Chlorotrimethylsilane (0.8 mL, 5.36 mmol) wasadded dropwise. The reaction mixture was warmed to r.t. and stirred for1 h. A solid of 5-bromomethyl-pyrazole-1-carboxylic acid tert-butylester (1.67 g, 6.4 mmol) was added and the reaction was heated to 45° C.overnight. The reaction mixture was cooled to rt, diluted with CH₂Cl₂,washed with aqueous NH₄Cl, dried with Na₂SO₄, and concentrated. Thecrude product was purified by combi-flash to give 682 mg of phosphinatein 55% yield.

To a solution of phosphinate (682 mg, 1.4 mmol) in CH₃CN (2 mL) at 0° C.was added iodotrimethylsilane (1.0 mL, 7 mmol). The reaction mixture waswarmed to r.t. and stirred for 30 minutes. An additional amount ofiodotrimethylsilane (1 mL, 7 mmol) was added and stirred for minutes.2,6-Lutidine (0.8 mL) and MeOH (1.6 mL) were added, stirred for 20minutes, concentrated in vacuo, and dried for 20 minutes to give amine.Coupling with intermediate VII gave phosphinic acid 85. ¹H NMR (300 MHz,CD₃OD):

8.30 (d, J=7.8 Hz, 1H), 8.17 (s, 1H) 7.76 (s, 2H), 7.61 (m, 4H), 7.61(s, 1H), 7.34 (d, J=9.3 Hz, 1H), 6.37 (s, 1H), 5.82 (m, 2H), 5.22 (dJ=17.7 Hz, 1H), 5.00 (d J=11.1 Hz, 1H), 4.68 (m, 3H), 4.49 (s, 1H), 4.16(m, 2H), 4.05 (m, 3H), 3.35 (m, 2H), 2.79 (m, 1H), 2.51 (m, 1H), 2.09(m, 1H), 1.63-1.48 (m, 8H), 1.34 (m, 6H), 1.02 (s, 9H). LC/MS=863.12(M⁺+1)

Example 86 Preparation of Compound 86

Step 1. To a solution of the phosphinate (structure shown above, 170 mg,0.44 mmol) in CH₃CN at 0° C. was added iodotrimethylsilane (0.³¹ mL,2.18 mmol). The reaction mixture was warmed to rt, stirred for 1 h, andcooled to 0° C. 2,6-lutidine (0.51 mL) was added followed by addition ofMeOH (0.5 mL) and warmed to rt. The mixture was concentrated and driedunder vacuum to give the desired amino phosphinic acid as crude product.

Step 2. The intermediate VII (142 mg, 0.22 mmol) and the aminophosphinic acid obtained from step 1 (0.44 mmol) were dissolved in DMF(2 mL). HATU (166 mg, 0.44 mmol) and NMM (0.07 mL, 0.65 mmol) were addedand the mixture was stirred at r.t. overnight. The reaction was dilutedwith CH₂Cl₂ and washed with 5% LiCl (2×). The organic layer was washedwith saturated NaHCO₃, dried with Na₂SO₄, and concentrated. The crudeproduct was purified by HPLC to give 83.2 mg of compound 86. ¹H NMR (300MHz, CD₃OD): d 8.28 (d, J=9.3 Hz, 1H), 8.17 (s, 1H), 7.96 (s, 1H), 7.77(s, 2H), 7.30 (dd, J=2.4, 9.0 Hz, 1H), 7.07 (s, 1H), 5.97 (m, 1H), 5.79(brs, 1H), 5.23 (d, J=17.7 Hz, 1H), 5.06 (d, J=11.7 Hz, 1H), 4.65 (m,2H), 4.46 (brs, 1H), 4.15 (m, 3H), 3.97 (s, 3H), 3.6 (d, 2H), 2.80 (m,1H), 2.45 (m, 1H), 2.12 (m, 1H), 1.4-1.7 (m, 10H), 1.34 (d, J=6.3 Hz,6H), 0.95-1.15 (brs, 9H); ³¹P (121.4 MHz, CD₃OD):

36.884; LC/MS=864 (M⁺+1).

Example 87 Preparation of Compound 87

A flask was charged with 1.1 mL (10.2 mmol) of freshly distilled2,5-dimethylthiazole and 25 mL of dry THF. To this mixture, 4.6 mL (2.8mmol) of 2.2 M nBuLi was then added dropwise and the reaction stirred at−78° C. for 30 min. The intermediate phosphonous acid IV (prepared from1.1 g of III, 3.4 mmol) was dissolved in 20 mL of dry THF and addeddropwise to the lithium anion solution of 2.5-Dimethylthiazole formed insitu at −78° C. After 30 min the reaction was quenched at −78° C. by theaddition of sat. NH₄Cl_((aq.)). The organic phase was diluted with EtOAcand washed with sat. NH₄Cl_((aq.)) and brine. The organic phase wasdried over MgSO₄. Concentration of the filtrate from vacuum filtrationremoval of the MgSO₄ yielded an orange oil from which product wasisolated by column chromatography (SiO₂, 3:1-Ethyl Acetate:Hexane) as aclear oil (220 mg, 15% over 2 steps).

¹H NMR (300 MHz, CDCl₃) d 7.33 (s, 5H), 6.64 (d, 1H), 5.80 (dt, J=9.9,17.1 Hz, 1H), 5.18 (b, 4H), 4.10 (m, 2H), 3.60 (m, 2H), 2.0 (m, 1H) 1.80(m, 2H), 1.20 (m, 3H). ³¹P NMR (121.4 MHz, CDCl₃) d 44.952, 41.135.LC/MS=421 (M⁺+1)

Deprotection and coupling as described before provide 87. (yield=65 mg,66%). ¹H NMR (300 MHz, CD₃OD) 8.28 (d, J=9.6 Hz, 1H), 8.27 (s, 1H), 7.77(s, 2H), 7.45 (s, 1H), 7.³¹ (dd, J=2.1, 9.3 Hz, 1H), 5.84 (br, 1H), 5.70(m, 1H), 5.12 (d, J=11.4 Hz, 1H), 4.83 (d, 1H) 4.69 (m, 2H), 4.51 (s,1H), 4.17 (m, 2H), 4.08 (s, 1H), 4.04 (s, 3H), 3.80 (m, 2H), 2.84 (dd,J=7.2, 14.1 Hz, 1H), 2.45 (m, 4H), 1.41-1.78 (m, 8H), 1.34 (d, J=6.3 Hz,6H), 1.04 (s, 9H). ³¹P NMR (121.4 MHz, CD₃OD) d 26.015. LC/MS=894 (M⁺+1)

Example 88 Preparation of Compound 88

A solution of the phosphonous acid IV (501.3 mg, 1.62 mmol), Hunig'sBase (680 μL, 3.90 mmol), and chlorotrimethylsilane (460 μL, 3.62 mmol)in CH₂Cl₂ (8 mL) was stirred at rt for 20 minutes. A solution of4-(chloromethyl)-2-methylthiazole (510 mg, 2.77 mmol),tetrabutylammonium iodide (598.4 mg, 1.620 mmol) and Hunig's Base (530μL, 3.04 mmol) in CH₂Cl₂ (1.5 mL) was added via a cannula at rt. Theresulting solution was stirred at 40° C. for 4.5 days and cooled to rt.The solution was concentrated and the residue dissolved in ethyl acetate(30 mL). The organic layer was washed with H₂O (×2) and the aqueouslayer extracted with ethyl acetate (30 mL). The organic fractions weredried (MgSO₄) and concentrated. The residue was purified with aCombiFlash column chromatography using hexane:ethyl acetate as eluent toobtain phosphinate (449 mg, 66%). ^(1H) NMR (300 MHz, CDCl₃): d 7.35 (s,2.15H), 7.33 (s, 2.85H), 7.03 (d, J=3.3 Hz, 0.43H), 6.94 (d, 3.9 Hz,0.57H), 6.72 (s, 0.57H), 6.60 (s, 0.43H), 6.04 (m, 1H), 5.71 (s, 1H),5.40-5.34 (d, J=17.1 Hz, 1H), 5.29 (s, 2H), 5.10 (m, 3H), 4.76-4.73 (d,J=10.2 Hz, 1H), 4.20 (m, 2H), 3.55 (m, 2H), 3.32 (m, 2H), 2.67 (s, 3H),2.27 (m, 2H), 1.71 (m, 4H). 1.23 (m, 3H), 1.13 (m, 1H), 0.93 (m, 1H).³¹P (121.4 MHz, CD₃OD):

48.382, 47.151, 44.628, 43.811

Example 89 Preparation of Compound 89

In a manner similar to example 7. (the other furan) Yield=230 mg (40%).¹H NMR (300 MHz, CDCl₃) 7.33 (s, 5H), 6.41 (d, 1H), 6.00 (m, 1H), 5.30(m, 1H), 5.08 (m, 3H), 4.05 (m, 2H), 2.96 (m, 2H), 2.08 (m, 1H)), 1.77(m, 1H), 1.46 (m, 1H), 1.21 (m, 3H).

³¹P NMR (121.4 MHz, CDCl₃) d 45.73, 42.55

LC/MS=390 (M⁺+1)

¹H NMR (300 MHz, CD₃OD) 8.27 (d, J=9.3 Hz, 1H), 8.19 (s, 1H), 7.75 (s,2H), 7.42 (m, 1H), 7.30 (m, 1H), 6.46 (s, 1H), 5.95 (m, 1H), 5.80 ((b,1H), 5.24 (d, J=17.1 Hz, 1H), 5.06 (d, J=10.2 Hz, 1H), 4.67 (m, 2H),4.45 (s, 1H), 4.17 (m, 2H), 4.11 (s, 1H), 4.04 (s, 3H), 3.06 (d, 15 Hz,2H), 2.77 (m, 1H), 2.45 (m, 1H), 2.07 (m, 1H), 1.41-1.78 (m, 8H), 1.34(d, J=6.3 Hz, 6H), 1.04 (s, 9H). ³¹P NMR (121.4 MHz, CD₃OD) d 41.17.LC/MS=863 (M⁺+1)

Example 90 Preparation of Compound 90

A flask was charged with 13.0 mL of dry ether and 2.95 mL of 2.2 M nBuLiin hexane. The mixture was cooled to −78° C. Freshly distilledbromothiazole 587 μl (6.5 mmol) was then added and the reaction stirredat −78° C. for 20 minutes. The intermediate phosphonous acid IV(prepared from 1.0 g of III, 3.1 mmol) was dissolved in 13.0 mL of THFand then added dropwise to the anion solution at −78° C. After 20 minthe reaction was quenched at −78° C. by the addition of sat.NH₄Cl_((aq.)). The organic phase was diluted with EtOAc and washed withsat. NH₄Cl_((aq.)) and brine. The organic phase was dried over MgSO₄.Concentration of the filtrate from vacuum filtration removal of theMgSO₄ yielded an orange oil from which product was isolated by columnchromatography (SiO₂, neat Ethyl Acetate) as a clear oil (450 mg, 37%over 2 steps).

¹H NMR (300 MHz, CDCl₃) d 8.07 (s, 1H), 7.667 (s, 1H), 7.33 (s, 5H),6.20-5.90 (m, 1H), 5.82 (s, 1H), 5.55 (d, J=38.1 Hz, 1H), 5.20 (m, 1H),5.06 (m, 3H), 4.24 (m, 2H), 2.05 (m, 1H), 1.96-1.70 (m, 2H), 1.52 (m,1H), 1.303 (m, 3H). ³¹P NMR (121.4 MHz, CDCl₃) d 28.845, 26.156.LC/MS=393 (M⁺+1), 415 (M⁺+Na)

A solution of phosphinate obtained above (300 mg, 0.77 mmol) in ACN (6.5mL) was cooled to 0° C. and TMSI (764 μL, 5.4 mmol) was added in adrop-wise fashion. The reaction was warmed to rt and stirred for anhour. The reaction was cooled back to 0° C. and 2,6-lutidine (897 μL,2.6 mmol) was added in a drop-wise fashion. This was followed by theaddition of Et₃N (2.7 mL, 19.3 mmol) and MeOH (8 mL). The reaction wasthen concentrated in vacuum and crude was used directly in the nextreaction.

A solution of dipeptide VII (150 mg, 0.23 mmol) in THF (4 mL) was cooledto −30° C. Et₃N (81 μL, 0.58 mmol) was added to this solution followedby ClCO₂Et (44 μL, 0.46 mmol). The reaction was stirred at a temperaturebetween −20° C. and −30° C. for 30 min.

A solution of crude from step 1 in CH₂Cl₂ (2 mL) was added in adrop-wise fashion at −30° C. and the reaction was warmed to rt andstirred for 2 hours. The reaction was quenched by the addition of sat.NH₄Cl_((aq.)). The organic phase was diluted with EtOAc and extractedwith sat. NH₄Cl_((aq.)), H₂O, and brine. The organic phase was thendried over Na₂SO₄, which was subsequently removed by vacuum filtration.The filtrate was concentrated in vacuum and the residue was dissolved inMeOH (1.5 mL). Compound 90 was isolated from this solution byreverse-phase HPLC as a yellow solid (82 mg, 41%). ¹H NMR (300 MHz,CD₃)D) d 8.25 (m, 2H), 8.20 (m, 2H), 8.02 (s, 1H), 7.75 (s, 2H), 7.39(d, J=8.7 Hz, 1H), 5.97 (b, 2H), 5.77 (b, 1H), 5.06 (d, J=11.4 Hz, 1H),4.63 (m, 2H), 4.44 (s, 1H), 4.17 (m, 2H), 4.08 (s, 1H), 4.04 (s, 3H),2.75 (b, 1H), 2.57 (b, 1H), 2.10 (b, 1H), 1.7-1.5 (b, 8H), 1.34 (d,J=6.3 Hz, 6H), 1.04 (s, 9H). ³¹P NMR (121.4 MHz, CD₃CN) d 18.28

LC/MS=866 (M⁺+1)

Example 91 Preparation of Compound 91

The[(1-Benzyloxycarbonylamino-2-vinyl-cyclopropyl)-ethoxy-phosphinoyl]-aceticacid from example 11 (290 mg, 0.79 mmol) was suspended in 4 mL of DMF.HATU (901 mg, 2.37 mmol), methylamine (133 mg, 1.97 mmol), followed byNMM (781 μl, 7.11 mmol) was added. After 2 hours, the reaction wasconcentrated and partitioned with EtOAc and H2O. The aqueous layer wasextracted 3× with EtOAc. The organic layer was dried over MgSO4,filtered and concentrated. The material, a brown oil (264 mg, 88%) wasused crude.

The residue was suspended in 1 mL of CH₃CN and cooled to 0° C.Iodotrimethylsilyl (TMSI) (187 μl, 1.³¹ mmol) was added and the solutionwas warmed to rt. After 45 minutes, the solution was cooled again to 0°C. and triethylamine (1 mL, 7.33 mmol) and 2 mL of MeOH. The solutionwas warmed to rt and stirred for an additional 20 minutes. The solutionwas concentrated, azeotroped 2× with toluene and put on high vacuum for30 minutes. Coupling with intermediate VII gave 91 as a yellow solid.^(1H) NMR (300 MHz, CD₃OD): d 8.23 (d, J=9.5 Hz, 1H), 8.20 (s, 1H), 7.79(s, 2H), 7.33 (d, J=8.8 Hz, 1H), 5.95 (m, 1H), 5.78 (s, 1H), 5.22 (d,J=9.6 Hz, 2H), 5.13 (d, J=9.0 Hz, 2H), 4.63 (m, 2H), 4.45 (bs, 1H), 4.20(m, 3H), 4.05 (s, 3H), 3.22 (m, 1H), 3.20 (d, 1H), 3.18 (s, 1H), 2.80(m, 1H), 2.78 (s, 3H), 2.45 (m, 1H), 2.15 (m, 1H), 1.62 (m, 2H), 1.50(m, 6H) 1.38 (d, 6H), 1.05 (s, 9H). ³¹P (121.4 MHz, CD₃OD):

36.642

Example 92 Preparation of Compound 92

Step 1. Dipeptide compound (Boc deprotected from intermediate XII) (2.86g, 4.27 mmol) and 2-tert-butoxycarbonylamino-3-hydroxy-butyric acid (958mg, 4.37 mmol) were dissolved in DMF (18 mL) and cooled to 0° C. TEA(1.09 mL, 8.54 mmol), HOBT (634 mg, 4.7 mmol), and EDCI (1.7 g) wereadded sequentially. The reaction mixture was stirred at 0° C. for 1 hand warmed to r.t. overnight. The reaction mixture was quenched with H₂Oand extracted with EtOAc (2×). The organic layer was washed with 5%LiCl, saturated NH₄Cl and brine, dried with Na₂SO₄, and concentrated.The crude product was purified by combi-flash to give 2.21 g oftripeptide in 62% yield. ³¹P NMR (121.4 MHz, CDCl₃) d 46.4, 43.9.LC/MS=836.0 (M⁺+1), 856.0 (M⁺+Na).

Step 2. Alcohol from step 1 (2.06 g, 2.5 mmol) and Pent-4-enoic acid(0.64 mL, 6.25 mmol) were dissolved in CH₂Cl₂ (18.75 mL)/DMF (6.25 mL).EDCI (1.8 g, 9.38 mmol) and DMAP (92 mg, 0.75 mmol) were addedsequentially. The reaction mixture was stirred at r.t. for 7 h andconcentrated. The reaction mixture was diluted with H₂O and extractedwith EtOAc (2×). The organic layer was washed with 5% LiCl and brine,dried with Na₂SO₄, and concentrated. The crude product was purified bycombi-flash to give 2.16 g of ester product in 96% yield. ³¹P NMR (121.4MHz, CDCl₃) d 44.5, 43.9, 43.2, 42.3. LC/MS=917.9 (M⁺+1), 856.0 (M⁺+Na).

Step 3. Ester (2.16 g, 2.36 mmol) was dissolved in CH₂Cl₂ (236 mL) anddegassed with N₂ for 20 minutes. Grubb's G1 (486 mg, 0.59 mmol) wasadded and degassed for an additional 20 minutes. The reaction mixturewas heated to 50° C. for 5.5 h and cooled to rt.Tris(hydroxymethyl)phosphine (3.66 g, 29.5 mmol) was added followed byaddition of TEA (8.2 mL, 59 mmol) and H₂O (20 mL). The reaction mixturewas heated to 50° C. for 5 h and then r.t. overnight. The two layerswere separated. The organic layer was washed with 0.5 N HCl and brine,dried with Na₂SO₄, and concentrated. The crude product was purified bycombi-flash to give 1.48 g of cyclized compound in 71% yield. ³¹P NMR(121.4 MHz, CDCl₃) d 44.4, 43.1. LC/MS=888.1 (M⁺+1), 909.9 (M⁺+Na)

Step 4. To a solution of cyclic olefin (1.48 g, 1.67 mmol) in CH₂Cl₂ (10mL) was added 4 N HCl in 1,4-dioxane (6.26 mL, 25.05 mmol). The reactionmixture was stirred at r.t. for 3.5 h, concentrated, dried under vacuumovernight, and then dissolved in THF (14.3 mL)/H₂O (2.4 mL). CompoundCarbonic acid cyclopentyl ester 2,5-dioxo-pyrrolidin-1-yl ester (398 mg,1.75 mmol) and TEA (0.7 mL, 5.01 mmol) were added. The reaction wasstirred at r.t. for 2 h and additional Carbonic acid cyclopentyl ester2,5-dioxo-pyrrolidin-1-yl ester (38 mg) was added. The reaction wasstirred for 2 h. The reaction was quenched by adding 0.5 N HCl anddiluted with EtOAc. The two layers were separated. The organic layerswere washed with 0.5 N HCl and brine, dried with Na₂SO₄, andconcentrated. The crude product was purified by combi-flash to give 1.45g of cyclopentyl carbamate in 96% yield. ³¹P NMR (121.4 MHz, CDCl₃) d44.4, 43.1. LC/MS=902.0 (M⁺+1).

Step 5. A solution of cyclopentyl carbamate (1.4 g, 1.55 mmol) and8-chloro-2-(2-isopropylamino-thiazol-4-yl)-7-methoxy-quinolin-4-ol (542mg, 1.55 mmol) in NMP (15 mL) was treated with Cs₂CO₃ (1.26 g, 3.88mmol). The reaction mixture was heated to 63° C. for 5 h and then cooledto rt. The reaction was diluted with EtOAc and washed with NaHCO₃. Theorganic layer was washed with 5% LiCl and brine, dried with Na₂SO₄, andconcentrated. The crude product was purified by combi-flash to give 1.18g of desired product in 75% yield.

Step 6. To a solution of product obtained above (1.18 g, 1.16 mmol) inCH₃CN (12 mL) at 0° C. was added 2,6-lutidine (1.35 mL, 11.6 mmol) andiodotrimethylsilane (1.66 mL, 11.6 mmol). The reaction mixture wasstirred at r.t. for 3 h and cooled to 0° C. 2,6-lutidine (0.27 mL, 2.32mmol) was added followed by addition of MeOH (5 mL) and warmed to rt for10 minutes. The mixture was concentrated, dried under vacuum. The crudeproduct was purified by reverse phase combi-flash followed by HPLC togive 1.01 g of 92 in 88% yield. ¹H NMR (300 MHz, CD₃OD)

8.26 (m, 2H), 7.85 (s, 1H), 7.68 (d, J=9.6 Hz, 1H), 7.26 (t, J=8.1 Hz,1H), 6.93 (t, J=7.8 Hz, 2H), 5.84 (m, 2H), 5.67 (t, J=10.8 Hz, 1H), 5.08(dd, J=6.3, 9.9 Hz, 1H), 4.75 (t, J=8.4 Hz, 1H), 4.68 (d, J=11.7 Hz,1H), 4.42 (d, J=9.9 Hz, 1H), 4.3 (m, 2H), 4.17 (s, 3H), 4.00 (quint.,J=6.6 Hz, 1H), 3.55 (t, J=15.3 Hz, 1H), 3.³¹ (t, J=15.3 Hz, 1H), 2.91(m, 2H), 2.6 (m, 1H), 2.46 (dd, J=5.4, 16.8 Hz, 1H), 2.30 (m, 1H), 2.20(m, 1H), 1.4-1.7 (brm, 10H), 1.37 (dd, J=2.1, 6.6 Hz, 6H), 1.25 (d,J=6.3 Hz, 3H), 1.04 (m, 1H). ³¹P NMR (121.4 MHz, CD₃OD) d 38.9.LC/MS=985.1 (M⁺+1)

Example 93 Preparation of Compound 93

Step 1. Ethyl acetimidate hydrochloride (1.23 g, 9.95 mmol) and2,2,2-trifluoroethylamine hydrochloride (1.35 g, 9.95 mmol) weredissolved in CH₂Cl₂ (32 mL)/H₂O (3.2 mL). K₂CO₃ (0.69 g, 4.98 mmol) wasadded and stirred for 30 minutes. The two layers were separated. Theaqueous layer was extracted with CH₂Cl₂ (2×10 mL). The combined organiclayer was dried with Na₂SO₄ and concentrated to give 1.48 g ofN-(2,2,2-Trifluoro-ethyl)-acetimidic acid ethyl ester as a light yellowliquid in 87% yield.

Step 2. The Boc protected tripeptide phosphinate was prepared in asimilar manner as described for example 58, which was deprotected as thefollowing. Tripeptide (500 mg, 0.54 mmol) was dissolved in CH₃CN (5 mL)and cooled to 0° C. Iodotrimethylsilane (0.77 mL) was added. Thereaction mixture was warmed to rt, stirred for 0.5 h, and cooled to 0°C. 2,6-lutidine (1.30 mL) was added followed by addition of MeOH (5 mL).The mixture was concentrated, co-evaporated with CH₂Cl₂ (2×), and driedin vacuo to give amino phosphinate as a 2,6-lutidine salt. The salt (80mg, 0.025 mmol) was dissolved in DMF (0.45 mL) and 0.1 N phosphatebuffer (0.9 mL). 2 N NaOH (86 μL) was added to adjust pH to 9. Asolution of N-(2,2,2-Trifluoro-ethyl)-acetimidic acid ethyl ester (150mg, 0.89 mmol) in DMF (0.1 mL) was added and stirred for 18 h. Thereaction mixture was filtered and the filtrate was purified by HPLC togive 8.8 mg of 93 as a yellow solid.

Example 94 Preparation of Compound 94

A solution of(1-benzyloxycarbonylamino-2-vinyl-cyclopropyl)-methyl-phosphinic acidethyl ester (100 mg, 0.308 mmol) in ACN (7.7 mL) was cooled to 0° C. andTMSI (220 μL, 1.54 mmol) was added in a drop-wise fashion. The reactionwas warmed to rt and stirred for an hour. The reaction was then cooledto 0° C. and additional TMSI (110 μL, 0.77 mmol) was added in adrop-wise fashion. The reaction was warmed to rt and stirred for 30 min.The reaction was cooled back to 0° C. and 2,6-lutidine (360 μL, 3.1mmol) was added in a drop-wise fashion. This was followed by theaddition of Et₃N (1 mL, 7.2 mmol) and MeOH (4 mL). The reaction was thenconcentrated in vacuum and crude was used directly in the next reaction.

A solution of VI (72 mg, 0.123 mmol) in THF (2 mL) was cooled to −30° C.Et₃N (34 μL, 0.246 mmol) was added to this solution followed by ClCO₂Et(18 μL, 0.185 mmol). The reaction was stirred at a temperature between−20° C. and −30° C. for 30 min. Additional Et₃N (34 μL, 0.246 mmol) andClCO₂Et (18 μL, 0.185 mmol) was added to the reaction. The reaction wasstirred for an additional 30 min at a temperature between −20° C. and−30° C. A solution of crude product from step 1 in CH₂Cl₂ (2 mL) wasadded in a drop-wise fashion at −30° C. and the reaction was warmed tort and stirred for 2 hours. The reaction was quenched by the addition ofsat. NH₄Cl_((aq.)). The organic phase was diluted with EtOAc andextracted with sat. NH₄Cl_((aq.)), H₂O, and brine. The organic phase wasthen dried over Na₂SO₄, which was subsequently removed by vacuumfiltration. The filtrate was concentrated in vacuum and the residue wasdissolved in MeOH (1.5 mL). Compound 94 was isolated from this solutionby reverse-phase HPLC as a yellow solid (35 mg, 38%). ¹H NMR (300 MHz,CD₃OD) d 8.25 (d, J=9.3 Hz, 1H), 8.16 (m, 2H), 7.68 (m, 3H), 7.49 (m1H), 7.39 (m 1H), 7.24 (dd, J=2.1, 9.3 Hz, 1H), 6.45 (m, 1H), 5.97 (m,2H), 5.69 (s, 1H), 5.26 (d, J=17.1 Hz, 1H), 5.08 (d, J=11.4 Hz, 1H),4.63 (m, 2H), 4.24 (m, 1H), 4.08 (m, 1H), 4.04 (s, 3H), 2.76 (dd, J=7.2,14.1 Hz, 1H), 2.43 (ddd, J=3.3, 10.5, 13.8 Hz, 1H), 1.42-1.78 (m, 8H),1.34 (d, J=6.3 Hz, 3H), 1.34 (m, 1H), 1.15 (m, 1H), 1.04 (s, 9H). ³¹PNMR (121.4 MHz, CD₃OD) d 41.2. LC/MS=733 (M⁺+1), 755 (M⁺+Na).

Example 95 Preparation of Compound 95

To a mixture of IX (12.38 g, 26.68 mmol) and2-Amino-7-methoxy-quinolin-4-ol (7.11 g, 37.35 mmol) in NMP (133 mL) wasadded Cs₂CO₃ (10.43 g, 32.01 mmol). The reaction mixture was heated to80° C. overnight and cooled to rt. The mixture was poured into brine(500 mL) and extracted with EtOAc (600 mL). The organic layer was washedwith saturated NaHCO₃ (300 mL), brine (200 mL), dried with Na₂SO₄, andconcentrated. The crude product was purified by combi-flash to give 4.02g of 4-(2-Amino-7-methoxy-quinolin-4-yloxy)-pyrrolidine-1,2-dicarboxylicacid 1-tert-butyl ester 2-methyl ester in 26% yield.

A mixture of ester (4.02 g, 9.62 mmol) and 3-methyl-1-nitrosooxy-butane(7.18 mL, 48.5 mmol) in HOAc (21 mL) was stirred at rt for 36 h, pouredinto H₂O (500 mL), and extracted with CH₂Cl₂ (2×150 mL). The aqueouslayer was diluted with brine (200 mL) and extracted with 10% MeOH/CH₂Cl₂(2×150 mL). The combined organic layers were dried with Na₂SO₄ andconcentrated. The crude product was purified by combi-flash to give 3.39g of4-(2-hydroxy-7-methoxy-quinolin-4-yloxy)-pyrrolidine-1,2-dicarboxylicacid 1-tert-butyl ester 2-methyl ester in 79% yield.

Crude4-(2-hydroxy-7-methoxy-quinolin-4-yloxy)-pyrrolidine-1,2-dicarboxylicacid 1-tert-butyl ester 2-methyl ester (3.18 g, 7.60 mmol) was dissolvedin POCl₃ (76 mL) and heated to 40° C. overnight. The reaction mixturewas concentrated in vacuo and dissolved in CH₂Cl₂ (40 mL). 4 N HCl in1,4-dioxane (40 mL) was added and stirred at r.t. for 1 h. The crudematerial was partitioned between H₂O and CH₂Cl₂ and pH was adjusted to11 with NaHCO₃ and 1 N NaOH. The aqueous layer was extracted with CH₂Cl₂(3×150 mL). The combined organic layer was dried with Na₂SO₄ andconcentrated. The crude product was purified by combi-flash to give 2.05g of4-(2-Chloro-7-methoxy-quinolin-4-yloxy)-pyrrolidine-1,2-dicarboxylicacid 1-tert-butyl ester 2-methyl ester.

Amine obtained above (562 mg, 1.67 mmol),2-Cyclopentyloxycarbonylamino-3,3-dim ethyl-butyric acid (489.8 mg, 2.01mmol) and HATU (1.27 g, 3.34 mmol) were dissolved in DMF (16 mL) andcooled to 0° C. NMM (0.74 mL, 6.73 mmol) was added. The reaction mixturewas warmed to r. t. and stirred overnight. The product was partitionedbetween H₂O (300 mL) and EtOAc (3×100 mL). The combined organic layerswere washed with H₂O (100 mL), NH₄Cl (100 mL), NaHCO₃ (100 mL), brine(100 mL), dried with Na₂SO₄, filtered, and concentrated. The crudeproduct was purified by combi-flash to give 691.6 mg of dipeptide methylester in 71% yield.

Ester (664 mg, 1.18 mmol) was dissolved in THF (4 mL), H₂O (4 mL), andMeOH (4 mL) and LiOH (142.2 mg, 5.94 mmol) was added. The reactionmixture was stirred at r. t. for 2 h. The mixture was diluted with H₂O(15 mL)/EtOAc (20 mL), acidified to pH=2 with 1.0 N HCl, and the twolayers were separated. The aqueous layer was extracted with EtOAc (3×50mL), dried with Na₂SO₄, filtered, and concentrated to give 661 mg ofacid.

Acid (9³¹ mg, 1.87 mmol),(1-amino-2-vinyl-cyclopropyl)-benzyl-phosphinic acid ethyl ester (548.1mg, 2.07 mmol) and HATU (1.42 g, 3.74 mmol) were dissolved in DMF (19mL) and cooled to 0° C. NMM (1.03 mL, 9.37 mmol) was added. The reactionmixture was warmed to r. t. and stirred overnight. The product waspartitioned between H₂O (300 mL) and EtOAc (3×100 mL). The combinedorganic layers were washed with H₂O (100 mL), NH₄Cl (100 mL), NaHCO₃(100 mL), brine (100 mL), dried with Na₂SO₄, filtered, and concentrated.The crude product was purified by combi-flash to give 1.21 g oftripeptide phosphinate in 81% yield.

Tripeptide phosphinate was dissolved in CH₃CN (1 mL) and cooled to 0° C.Iodotrimethylsilane (72 μL, 0.51 mmol) was added dropwise. The reactionmixture was stirred at 0° C. for 45 min and 2,6-lutidine (0.5 mL) wasadded. MeOH (0.5 mL) was added and the reaction mixture wasconcentrated. The crude product was purified by HPLC to give 61.5 mg of95 in 79% yield. ¹H NMR (300 MHz, CDCl₃) d: 8.10 (d, J=9.3 Hz, 1H),7.37-7.17 (m, 6H), 7.14 (dd, J=9.3, 2.1 Hz, 1H), 7.06 (s, 1H), 5.95 (dt,J=17.1, 9.3 Hz, 1H), 5.48 (bs, 1H), 5.32-5.21 (m, 1H), 5.11-5.03 (m,1H), 4.68-4.51 (m, 3H), 4.22 (s, 1H), 4.06-3.98 (m, 1H), 3.95 (s, 1H),3.35-3.23 (m, 2H), 2.73-2.62 (m, 1H), 2.41-2.28 (m, 1H), 2.17-2.04 (m,1H), 1.82-1.33 (m, 10H), 1.03 (s, 9H); ³¹P (121.4 MHz, CDCl₃) d: 47.5;LCMS (M⁺+1): 767.06.

Example 96 Preparation of Compound 96

A solution of chloroquinoline from example 95 (51.7 mg, 0.06 mmol) inDMF (0.43 mL) was treated with 4-fluorophenylboronic acid (13.5 mg, 0.10mmol) and tetrakistriphenylphosphine palladium (7.3 mg). A solution ofK₂CO₃ (9 mg, 0.06 mmol) in H₂O (0.22 mL) was added to the above mixture.The reaction mixture was heated at 100° C. in microwave for 1 h. Thedesired ester (46.7 mg) was obtained after HPLC purification.

Ester (44.8 mg, 0.05 mmol) was dissolved in CH₃CN (0.53 mL) and cooledto 0° C. Iodotrimethylsilane (37.5 μL, 0.27 mmol) was added dropwise.The reaction mixture was stirred at 0° C. for 1 h and 2,6-lutidine (0.3mL) was added. MeOH (0.3 mL) was added and the reaction mixture wasconcentrated. The crude product was purified by HPLC to give 17.7 mg of96 in 41% yield.

¹H NMR (300 MHz, CDCl₃) d: 8.37 (d, J=9.0 Hz, 1H), 8.20-8.12 (m, 2H),7.71-7.44 (m, 7H), 7.44-7.18 (m, 3H), 5.96 (dt, J=16.8, 10.2 Hz, 1H),5.83 (bs, 1H), 5.25 (d, J=16.8 Hz, 1H), 5.11-5.04 (m, 1H), 4.74-4.65 (m,2H), 4.51-4.42 (m, 1H), 4.22-4.09 (m, 2H), 4.05 (s, 3H), 3.26 (d, J=15.6Hz, 2H), 2.87-2.76 (m, 1H), 2.54-2.41 (m, 1H), 2.16-2.03 (m, 1H),1.71-1.28 (m, 10H), 1.02 (s, 9H

³¹P (121.4 MHz, CDCl₃) d: 42.3, 32.6; LCMS (M+1): 827.0

Example 97 Preparation of Compound 97

A solution of chloroquinoline from example 95 (78.2 mg, 0.10 mmol) inDMF (0.65 mL) was treated with 2-fluorophenylboronic acid (20.1 mg, 0.15mmol) and tetrakistriphenylphosphine palladium (6.0 mg). A solution ofK₂CO₃ (13.6 mg, 0.10 mmol) in H₂O (0.33 mL) was added to the abovemixture. The reaction mixture was heated at 100° C. in microwave for 1h. The desired ester (73.3 mg) was obtained after HPLC purification.

The phosphinate ester obtained above (73.3 mg, 0.09 mmol) was dissolvedin CH₃CN (0.85 mL) and cooled to 0° C. Iodotrimethylsilane (61 μL, 0.43mmol) was added dropwise. The reaction mixture was stirred at 0° C. for1 h and 2,6-lutidine (0.3 mL) was added. MeOH (0.3 mL) was added and thereaction mixture was concentrated. The crude product was purified byHPLC to give 97. LCMS (M⁺+1): 827.15

Example 98 Preparation of Compound 98

A solution of chloroquinoline from example 95 (78.0 mg, 0.10 mmol) inDMF (0.98 mL) was treated with 3-fluorophenylboronic acid (21.9 mg, 0.16mmol) and tetrakistriphenylphosphine palladium (6.4 mg). A solution ofK₂CO₃ (13.5 mg, 0.1 mmol) in H₂O (0.3 mL) was added to the abovemixture. The reaction mixture was heated at 100° C. in microwave for 1h. The crude product was purified by HPLC to give 41 mg of ester in 49%yield.

Ester (41 mg, 0.05 mmol) was dissolved in CH₃CN (0.48 mL) and cooled to0° C. Iodotrimethylsilane (34 μL, 0.24 mmol) was added dropwise. Thereaction mixture was stirred at 0° C. for 1 h and 2,6-lutidine (0.3 mL)was added. MeOH (0.3 mL) was added and the reaction mixture wasconcentrated. The crude product was purified by HPLC to give 23 mg ofacid 98. LCMS (M+1): 827.13.

Example 99 Preparation of Compound 99

A solution of chloroquinoline from example 95 (78.4 mg, 0.10 mmol) inDMF (0.98 mL) was treated with 4-methoxyphenylboronic acid (23 mg, 0.15mmol) and tetrakistriphenylphosphine palladium (5.9 mg). A solution ofK₂CO₃ (13.5 mg, 0.1 mmol) in H₂O (0.3 mL) was added to the abovemixture. The reaction mixture was heated at 100° C. in microwave for 1h. The crude product was purified by HPLC to give 43.8 mg of ester in51% yield.

Ester (43.8 mg, 0.05 mmol) was dissolved in CH₃CN (0.5 mL) and cooled to0° C. Iodotrimethylsilane (36.5 μL, 0.26 mmol) was added dropwise. Thereaction mixture was stirred at 0° C. for 1 h and 2,6-lutidine (0.3 mL)was added. MeOH (0.3 mL) was added and the reaction mixture wasconcentrated. The crude product was purified by HPLC to give 29 mg ofacid 99.

Example 100 Preparation of Compound 100

A solution of chloroquinoline from example 95 (79.9 mg, 0.10 mmol) inDMF (0.98 mL) was treated with 2-methoxyphenylboronic acid (24.4 mg,0.16 mmol) and tetrakistriphenylphosphine palladium (5.9 mg). A solutionof K₂CO₃ (13.7 mg, 0.1 mmol) in H₂O (0.33 mL) was added to the abovemixture. The reaction mixture was heated at 100° C. in microwave for 1h. The crude product was purified by HPLC to give 29.9 mg of ester in36% yield.

Ester (29.9 mg, 0.03 mmol) was dissolved in CH₃CN (0.35 mL) and cooledto 0° C. Iodotrimethylsilane (25 μL, 0.18 mmol) was added dropwise. Thereaction mixture was stirred at 0° C. for 1 h and 2,6-lutidine (0.3 mL)was added. MeOH (0.3 mL) was added and the reaction mixture wasconcentrated. The crude product was purified by HPLC to give 19 mg ofacid 100.

Example 101 Preparation of Compound 101

Dipeptides in examples 101-103 are known in prior literature. Each wascoupled to the benzyl phosphinate P1 by the same method used in example35. Prep HPLC afforded (Example 101) (22 mg, 24%). ^(1H) NMR (300 MHz,CD₃OD): d 8.14 (d, J=8.8 Hz, 1H), 7.90 (d, J=6.0 Hz, 1H), 7.30 (m, 8H),5.91 (m, 1H), 5.84 (bs, 1H), 5.21 (m, 1H), 5.05 (m, 1H) 4.63 (m, 2H),4.50 (m, 1H), 4.03 (m, 1H), 3.94 (s, 3H), 3.32 (m, 2H), 2.65 (m, 1H),2.33 (m, 1H), 2.11 (m, 1H), 1.60 (m, 10H) 1.03 (s, 9H). ³¹P (121.4 MHz,CD₃OD):

42.175. LC (6 minute run, r.t.=4.15 min) MS (733.7, M+1)

Example 102 Preparation of Compound 102

Prep HPLC afforded (Example 102) (29 mg, 26%). ^(1H) NMR (300 MHz,CD₃OD): d 8.07 (d, J=9.2 Hz, 1H), 7.30 (m, 8H), 7.00 (d, J=8.9 Hz, 1H)6.82 (d, J=6.1 Hz, 1H) 5.92 (m, 1H), 5.30 (m, 2H), 5.05 (m, 1H) 4.83(bs, 1H) 4.58 (m, 1H), 4.42 (m, 1H), 4.03 (m, 1H), 3.90 (s, 3H), 3.32(m, 2H), 2.65 (m, 1H), 2.28 (m, 1H), 2.06 (m, 1H), 1.60 (m, 10H) 1.03(s, 9H). ³¹P (121.4 MHz, CD₃OD):

42.159. LC (6 minute run, r.t.=4.32 min) MS (732.7, M+1)

Example 103 Preparation of Compound 103

Prep HPLC afforded (Example 103) (15 mg, 19%). ^(1H) NMR (300 MHz,CD₃OD): d 8.18 (d, J=8.0 Hz, 1H), 7.80 (d, J=8.2 Hz, 1H), 7.30 (m, 8H),6.97 (d, J=7.6 Hz, 1H) 5.92 (m, 1H), 5.30 (m, 2H), 5.05 (m, 1H) 4.83(bs, 1H) 4.56 (m, 1H), 4.45 (m, 1H), 4.05 (m, 1H), 3.32 (m, 2H), 2.67(m, 1H), 2.28 (m, 1H), 2.07 (m, 1H), 1.60 (m, 10H) 1.03 (s, 9H). ³¹P(121.4 MHz, CD₃OD):

42.125

LC (6 minute run, r.t.=4.37 min) MS (702.7, M+1)

Example 104 Preparation of Compound 104

Compound 94 (100 mg, 0.14 mmol) was dissolved in pyridine (3 mL)followed by the addition of phenol (129 mg, 1.37 mmol) and the solutionwas heated at 60° C. for 10 min. To the heated solution was addeddicyclohexylcarbodiimide (169 mgs, 0.82 mmol) and the reaction mixturewas further heated for 3 h. The reaction mixture was then cooled to rtand the solvents were removed under reduced pressure. The reactionmixture was diluted with EtOAc and the solids were filtered. Solvent wasremoved under reduced pressure and the crude product was purified oncombi-flash EtOAc/Hex to afford 23 mg of phosphinate prodrug in 21%yield. ¹H NMR (300 MHz, CDCl₃): d 8.07-7.99 (m, 2H), 7.55-7.00 (m, 11H),5.89-5.83 (m, 1H), 5.45-4.91 (m, 4H), 4.33-3.96 (m, 5H), 2.56 (m, 2H),1.97-0.90 (m, 28H). ³¹P (121.4 MHz, CDCl₃): 51.6 (s, ³¹P), 48.86 (s,³¹P); LC/MS: M+Na=8³¹.

Example 105 Preparation of Compound 105

Step 1. The 8-chloro-4-hydroxy-7-methoxy-quinoline-2-carboxylic acid(500 mg, 1.97 mmol) and 2.0 M dimethylamine in THF (2 mL, 3.94 mmol)were dissolved in DMF (20 mL). HATU (1.5 g, 3.94 mmol) and NMM (697 mg,6.89 mmol) were added and the mixture was stirred at r.t. overnight. Thereaction was diluted with EtOAc and acidified with 1 N HCl. The twolayers were separated. The organic layer was washed with 2% LiCl,saturated NaHCO₃ and brine, dried with Na₂SO₄, and concentrated to give8-chloro-4-hydroxy-7-methoxy-quinoline-2-carboxylic acid dimethylamide.

A solution of brosylate example 58 (100 mg, 0.11 mmol) and phenolobtained above (35 mg, 0.11 mmol) in NMP (5 mL) was treated with Cs₂CO₃(76 mg, 0.22 mmol). The reaction mixture was heated to 80° C. for 3 hand cooled to rt. The reaction was diluted with EtOAc and washed withH₂O. The aqueous layer was brought to pH=4 with 1 N HCl and extractedwith 5% MeOH/EtOAc (2×). The combined organic layers were dried withNa₂SO₄ and concentrated to give 58 mg of phosphinate.

To a solution of phosphinate (58 mg, 0.06 mmol) in CH₃CN (0.5 mL) at 0°C. was added iodotrimethylsilane (0.05 mL, 0.32 mmol). The reactionmixture was warmed to r.t. and stirred for 30 minutes and then cooled to0° C. 2,6-Lutidine (0.32 mL) and MeOH (0.5 mL) were added and stirredfor 10 minutes. The solvent was concentrated and the crude product waspurified by HPLC to give acid 105. ¹H NMR (300 MHz, CD₃OD):

8.20 (d, J=9.3 Hz, 1H), 7.48 (d, J=8.7 Hz, 1H), 7.28 (m, 1H), 7.14 (s,1H), 6.95 (m, 2H), 6.00 (m, 1H), 5.52 (m, 1H), 5.38 (m, 1H), 5.12 (m,1H), 4.62 (m, 2H), 4.³¹ (bs, 1H), 4.17 (s, 3H), 4.06 (m, 3H), 3.20 (m,6H), 2.74 (m, 1H), 2.42 (m, 1H), 2.20 (m, 1H), 1.70-1.40 (m, 8H), 1.30(m, 2H), 1.01 (s, 9H). LC/MS=874.13 (M⁺+1), 896.27 (M⁺+Na)

Example 106 Preparation of Compound 106

8-Chloro-4-hydroxy-7-methoxy-quinoline-2-carboxylic acid (115 mg, 0.45mmol) and ammoniumchloride (36 mg, 0.68 mmol) were dissolved in DMF (4mL). HATU (342 mg, 0.9 mmol) and NMM (159 mg, 1.58 mmol) were added andthe mixture was stirred at r.t. overnight. An additional amount ofammoniumchloride (72 mg, 13.5 mmol) was added and heated to 53° C. for18 h. The reaction mixture was cooled to r.t. and concentrated to give8-chloro-4-hydroxy-7-methoxy-quinoline-2-carboxylic acid amide as ayellow solid.

A solution of brosylate example 58 (380 mg, 0.44 mmol) and8-chloro-4-hydroxy-7-methoxy-quinoline-2-carboxyli c acid amide (100 mg,0.4 mmol) in NMP (3 mL) was treated with Cs₂CO₃ (287 mg, 0.88 mmol). Thereaction mixture was heated to 80° C. overnight and then cooled to rt.The reaction mixture was diluted with EtOAc and washed with H₂O. Theaqueous layer was brought to pH=4 with 1 N HCl and extracted with EtOAc(2×). The combined organic layers were dried with Na₂SO₄ andconcentrated. 334 mg of crude product was obtained.

To a solution of crude product obtained above (78 mg, 0.09 mmol) inCH₃CN (0.5 mL) at 0° C. was added iodotrimethylsilane (89 mg, 0.45mmol). The reaction mixture was stirred at 0° C. for 20 minutes.2,6-Lutidine (0.06 mL) and MeOH (0.5 mL) were added, stirred for 20minutes, concentrated in vacuo, and dried for 20 minutes to give acid,which was treated with TFAA to provide 106. ¹H NMR (300 MHz, CD₃OD):

8.21 (d, J=9.6 Hz, 1H), 7.56 (d, J=9.6 Hz, 2H), 7.44 (s, 1H), 7.28 (m,1H), 6.96 (m, 2H), 5.96 (m, 1H), 5.54 (s, 1H), 5.30 (d, J=17.1 Hz, 1H),5.11 (d, J=12.0 Hz, 1H), 4.69-4.56 (m, 2H), 4.36 (bs, 1H), 4.17-4.00 (m,6H), 3.38 (m, 2H), 2.74 (m, 1H), 2.45 (m, 1H), 2.20 (m, 1H), 1.67-1.54(m, 8H), 1.47 (m, 2H), 1.02 (s, 9H). ³¹P (121.4 MHz, CDCl₃): d 41.479.LC/MS=874.13 (M⁺+1), 896.27 (M⁺+Na)

Example 107 Preparation of Compound 107

A solution of the acid VI (160 mg, 0.27 mmol), HATU (256 mg, 0.67 mmol),acid (example 9) (80 mg, 0.54 mmol), and NMM (148 μl, 1.35 mmol) stirredin DMF (1 mL) overnight at rt. The solution was concentrated andpurified with a Gilson HPLC to obtain 107 (25.3 mg, 13%) as a whitesolid. ^(1H) NMR (300 MHz, CD₃OD): d 8.39 (d, J=9.0 Hz, 1H), 8.10 (d,J=9.3 Hz, 2H), 7.79 (m, 2H), 7.63 (s, 1H), 7.58 (s, 1H), 7.39 (d, J=8.8Hz, 1H), 5.87 (m, 2H), 5.30 (d, J=9.6 Hz, 1H), 5.18 (d, J=9.0 Hz, 1H)4.78 (m, 3H), 4.20 (s, 1H), 4.05 (s, 3H), 2.80 (m, 1H), 2.50 (m, 1H),2.10 (m, 1H), 1.62 (m, 2H) 1.40 (m, 2H) 1.05 (s, 9H). ³¹P (121.4 MHz,CD₃OD):

23.135

Example 108 Preparation of Compound 108

The P1 phosphinate amine was prepared as described in example 2 andcoupled with VI. ¹H NMR (300 MHz, CD₃OD)

8.36 (d, J=9.3 Hz, 1H), 8.1 (m, 2H), 7.76 (m, 3H), 7.65 (s, 1H), 7.55(d, J=2.1 Hz, 1H), 7.38 (dd, J=2.4, 9.3 Hz, 1H), 5.96 (dt, J=9.9, 17.1Hz, 1H), 5.85 (s, 1H), 5.26 (d, J=16.8 Hz, 1H), 5.08 (d, J=12 Hz, 1H),4.66 (m, 2H), 4.46 (s, 1H), 4.16 (s, 1H), 4.08 (m, 1H), 4.06 (s, 3H),2.78 (dd, J=6.6, 14.1 Hz, 1H), 2.43 (ddd, J=3.9, 10.2, 14.1 Hz, 1H),2.08 (m, 1H), 1.83 (m, 2H), 1.39-1.65 (brm, 10H), 1.14 (dt, J=7.8, 18.3Hz, 3H), 1.04 (s, 9H).

³¹P NMR (121.4 MHz, CD₃OD) d 50.6

LC/MS=746.8 (M⁺+1)

Example 109 Preparation of Compound 109

The P1 phosphinate amine was prepared as described in example 10 andcoupled with VI. ^(1H) NMR (300 MHz, CD₃OD): d 8.38 (d, J=9.6 Hz, 1H),8.11 (m, 2H), 7.76 (m, 3H), 7.65 (m 1H), 7.55 (m 1H), 7.24 (dd, J=2.1,9.6 Hz, 1H), 6.02 (m, 1H), 5.81 (m, 1H), 5.22 (d, J=9.6 Hz, 1H), 5.09(d, J=9.0 Hz, 1H), 4.63 (m, 2H), 4.45 (bs, 1H), 4.20 (m, 1H), 4.07 (m,4H), 2.80 (m, 1H), 2.41 (m, 1H), 2.10 (m, 1H), 1.89-1.33 (m, 13H), 1.05(m, 12H). ³¹P (121.4 MHz, CD₃OD):

48.663

LC/MS=761 (M⁺+1)

Example 110 Preparation of Compound 110

The acid VI (82 mg, 0.14 mmol) was suspended in 1 mL of DMF. HATU (133mg, 0.35 mmol),(1-Amino-2-vinyl-cyclopropyl)-(2-hydroxy-ethyl)-phosphinic acid (example24) (53 mg, 0.28 mmol), followed by NMM (77 μl, 0.70 mmol) was added.The solution stirred overnight at rt. The mixture was purified viaGilson HPLC to obtain 110 (28.3 mg, 27%) as a white solid. ^(1H) NMR(300 MHz, CD₃OD): d 8.39 (d, J=9.0 Hz, 1H), 8.10 (d, J=9.3 Hz, 2H), 7.79(m, 2H), 7.63 (s, 1H), 7.58 (s, 1H), 7.39 (d, J=8.8 Hz, 1H), 5.87 (m,2H), 5.30 (d, J=9.6 Hz, 1H), 5.18 (d, J=9.0 Hz, 1H) 4.78 (m, 3H), 4.20(s, 1H), 4.05 (s, 3H), 3.80 (m, 1H), 2.80 (m, 1H), 2.50 (m, 1H), 2.20(m, 1H), 2.10 (m, 1H), 1.62 (m, 2H) 1.40 (m, 2H) 1.20 (d, 3H), 1.05 (s,9H). ³¹P (121.4 MHz, CD₃OD):

44.493

Example 111 Preparation of Compound 111

A solution of the acid VI (100 mg, 0.17 mmol), HATU (161 mg, 0.42 mmol),amine example x (65 mg, 0.34 mmol), and NMM (93 μl, 0.85 mmol) stirredin DMF (1 mL) overnight at rt. The solution was concentrated andpurified with a Gilson HPLC to obtain 111 (97 mg, 75%) as a white solid.^(1H) NMR (300 MHz, CD₃OD): d 8.39 (d, J=9.0 Hz, 1H), 8.10 (d, J=9.3 Hz,2H), 7.79 (m, 2H), 7.63 (s, 1H), 7.58 (s, 1H), 7.39 (d, J=8.8 Hz, 1H),5.87 (m, 2H), 5.30 (d, J=9.6 Hz, 1H), 5.18 (d, J=9.0 Hz, 1H) 4.78 (m,3H), 4.20 (s, 1H), 4.05 (s, 3H), 2.80 (m, 1H), 2.50 (m, 1H), 2.10 (m,1H), 1.62 (m, 2H) 1.40 (m, 2H) 1.05 (s, 9H). ³¹P (121.4 MHz, CD₃OD):

37.043

Example 112 Preparation of Compound 112

The P1 phosphinate amine was prepared as described in example 17 andcoupled with VI. ^(1H) NMR (300 MHz, CD₃CN): d 8.25 (d, J=9.0 Hz, 1H),8.06 (min, 2H), 7.80 (m 1H), 7.63 (m 3H), 7.³¹ (m, 2H), 6.50 (m, 1H),5.89 (m, 2H), 5.64 (bs, 1H), 5.04 (d, J=16.5 Hz, 1H), 4.91 (d, J=10.5Hz, 1H), 4.62 (m, 3H), 4.20 (m, 1H), 3.98 (min, 4H), 2.67 (m, 1H), 2.39(m, 1H), 1.90 (m, 1H), 1.89-1.23 (m, 12H), 1.05 (s, 9H). ³¹P (121.4 MHz,CD₃CN):

33.463. LC/MS=759 (M⁺+1)

Example 113 Preparation of Compound 113

The P1 phosphinate amine was prepared as described in example 18 andcoupled with VI. ^(1H) NMR (300 MHz, CD₃OD): d 8.36 (d, J=9.6 Hz, 1H),8.08 (m, 2H), 7.76 (m, 3H), 7.65 (m 1H), 7.55 (m 1H), 7.39 (dd, J=2.1,9.6 Hz, 1H), 6.63-6.41 (m, 1H), 5.96 (m, 2H), 5.81 (bs, 1H), 5.25 (d,J=16.8 Hz, 1H), 5.07 (d, J=12.0 Hz, 1H), 4.65 (m, 2H), 4.44 (bs, 1H),4.15 (m, 1H), 4.04 (m, 4H), 2.77 (m, 1H), 2.43 (m, 1H), 2.10 (m, 3H),1.69-1.33 (m, 8H), 1.05 (s, 9H). ³¹P (121.4 MHz, CD₃OD):

33.412 LC/MS=759 (M⁺+1)

Example 114 Preparation of Compound 114

^(1H) NMR (300 MHz, CD₃OD): d 8.37 (d, J=9.0 Hz, 1H), 8.09 (m, 2H), 7.77(m, 3H), 7.67 (m 1H), 7.53 (m 1H), 7.38 (m, 1H), 5.98 (m, 1H), 5.84 (bs,1H), 5.24 (d, J=16.8 Hz, 1H), 5.07 (d, J=10.2 Hz, 1H), 4.70 (m, 2H),4.46 (bs, 1H), 4.17 (m, 1H), 4.06 (m, 4H), 2.80 (m, 1H), 2.54 (m, 1H),2.21 (m, 1H), 1.98 (m, 3H), 1.69-1.33 (m, 8H), 1.05 (s, 9H). ³¹P (121.4MHz, CD₃OD):

10.616. LC/MS=757 (M⁺+1)

Example 115 Preparation of Compound 115

The P1 phosphinate amine was prepared as described in example 35 andcoupled with VI. ^(1H) NMR (300 MHz, CD₃OD): d 8.38 (d, J=9.3 Hz, 1H),8.10 (m, 2H), 7.77 (m, 3H), 7.65 (m 1H), 7.55 (m 1H), 7.24 (m, 1H), 5.98(m, 1H), 5.87 (bs, 1H), 5.27 (d, J=17.1 Hz, 1H), 5.08 (d, J=10.5 Hz,1H), 4.72 (m, 2H), 4.46 (bs, 1H), 4.17 (m, 1H), 4.07 (m, 4H), 2.83 (m,1H), 2.48 (m, 1H), 2.09 (m, 1H), 1.97 (m, 3H), 1.69-1.33 (m, 10H), 1.05(s, 9H). ³¹P (121.4 MHz, CD₃OD):

40.676. LC/MS=809 (M⁺+1)

Example 116 Preparation of Compound 116

The P1 phosphinate amine was prepared as described in example 83 andcoupled with VI. ¹H NMR (300 MHz, CD₃OD) d 8.39 (d, J=9.6 Hz, 1H), 8.10(dd, J=1.2 Hz, 5.4 Hz, 2H), 7.77 (m, 5H), 7.68 (s, 1H), 7.55 (s, 1H),7.38 (m, 2H), 6.30 (m, 2H), 5.89 (m, 1H), 5.82 (s, 1H), 5.20 (d, J=17.1Hz, 1H), 5.07 (d, J=8 Hz, 1H), 4.75 (m, 2H), 4.51 (b, 1H), 4.17 (s, 1H),4.07 (s, 3H), 3.35 (m, 2H), 2.78 (m, 1H), 2.50 (m, 1H), 2.10 (m, 1H),1.41-1.78 (m, 8H), 1.04 (s, 9H). ³¹P NMR (121.4 MHz, CD₃OD) d 39.32.

LC/MS=799 (M⁺+1).

Example 117 Preparation of Compound 117

The P1 phosphinate amine was prepared as described in example 87 andcoupled with VI. ¹H NMR (300 MHz, CD₃OD) d 8.39 (d, J=9.6 Hz, 1H), 8.10(dd, J=1.2 Hz, 5.4 Hz, 2H), 7.77 (m, 5H), 7.68 (s, 1H), 7.55 (d, J=2.1Hz, 1H), 7.42 (m, 2H), 5.89 (br, 1H), 5.75 (m, 1H), 5.10 (d, J=17.1 Hz,1H), 4.80 (d, 1H) 4.69 (m, 2H), 4.51 (b, 1H), 4.17 (s, 1H), 4.07 (s,3H), 3.72 (m, 2H), 2.78 (m, 1H), 2.50 (m, 4H), 2.17 (m, 1H), 1.41-1.78(m, 8H), 1.04 (s, 9H). ³¹P NMR (121.4 MHz, CD₃OD) d 24.93.

LC/MS=830 (M⁺+1)

Example 118 Preparation of Compound 118

The P1 phosphinate amine was prepared as described in example 88 andcoupled with VI. ^(1H) NMR (300 MHz, CD₃OD): d 8.37 (d, 1H, J=9.3 Hz),8.05-8.17 (m, 2H), 7.70-7.83 (m, 3H), 7.68 (s, 1H), 7.50-7.62 (m, 2H),7.35 (dd, 1H, J=9.3 and 2.1 Hz), 5.89 (br, 1H), 5.72 (dt, 1H, J=17.1 and9.9 Hz), 5.09 (d, 1H, J=17.1 Hz), 4.70-5.04 (m, 4H), 4.51 (br, 1H), 4.21(br, 1H), 4.04-4.18 (m, 1H), 4.05 (s, 3H), 3.36-3.50 (m, 2H), 2.82-2.94(m, 1H), 2.80 (s, 3H), 2.50-2.65 (m, 1H), 2.09 (br m, 1H), 1.32-1.80 (m,9H), 1.05 (s, 9H). ³¹P (121.4 MHz, CD₃OD): d 33.449. LC/MS=830 (M⁺+1)

Example 119 Preparation of Compound 119

The acid VI (85 mg, 0.14 mmol) was suspended in 1 mL of DMF. HATU (133mg, 0.35 mmol), (1-amino-2-vinyl-cyclopropyl)-carbamoylmethyl-phosphinicacid (59 mg, 0.29 mmol), followed by NMM (77 μl, 0.70 mmol) was added.The solution stirred overnight at rt. The mixture was purified viaGilson HPLC to obtain 119 (30 mg, 27%) as a white solid. ^(1H) NMR (300MHz, CD₃OD): d 8.39 (d, J=9.0 Hz, 1H), 8.10 (d, J=9.3 Hz, 2H), 7.79 (m,2H), 7.63 (s, 1H), 7.58 (s, 1H), 7.39 (d, J=8.8 Hz, 1H), 5.87 (m, 2H),5.83 (s, 1H), 5.22 (d, J=9.6 Hz, 1H), 5.13 (d, J=9.0 Hz, 1H), 4.78 (m,2H), 4.50 (s, 1H), 4.20 (s, 1H), 4.05 (s, 3H), 3.15 (m, 1H), 2.80 (m,1H), 2.50 (m, 1H), 2.10 (m, 1H), 1.62 (m, 4H), 1.40 (m, 2H), 1.05 (s,9H). ³¹P (121.4 MHz, CD₃OD):

36.428

Example 120 Preparation of Compound 120

The acid VI (76 mg, 0.13 mmol) was suspended in 1 mL of DMF. HATU (123mg, 0.32 mmol),(1-amino-2-vinyl-cyclopropyl)-methylcarbamoylmethyl-phosphinic acid (56mg, 0.26 mmol), followed by NMM (71 μl, 0.65 mmol) was added. Thesolution stirred overnight at rt. The mixture was purified via GilsonHPLC to obtain 120 (93 mg, 91%) as a white solid. ^(1H) NMR (300 MHz,CD₃OD): d 8.39 (d, J=9.0 Hz, 1H), 8.10 (d, J=9.3 Hz, 2H), 7.79 (m, 2H),7.63 (s, 1H), 7.58 (s, 1H), 7.39 (d, J=8.8 Hz, 1H), 5.87 (m, 2H), 5.30(d, J=9.6 Hz, 1H), 5.18 (d, J=9.0 Hz, 1H) 4.78 (m, 3H), 4.20 (s, 1H),4.05 (s, 3H), 3.30 (m, 1H), 3.20 (d, 1H), 3.18 (m, 3H), 2.87 (s, 3H),2.80 (m, 1H), 2.50 (m, 1H), 2.10 (m, 1H), 1.62 (m, 2H) 1.40 (m, 2H) 1.20(d, 3H), 1.05 (s, 9H). ³¹P (121.4 MHz, CD₃OD):

37.802

Example 121 Preparation of Compound 121

The acid VI (100 mg, 0.17 mmol) was suspended in 1 mL of DMF. HATU (161mg, 0.42 mmol),(1-amino-2-vinyl-cyclopropyl)-dimethylcarbamoylmethyl-phosphinic acid(78 mg, 0.34 mmol), followed by NMM (93 μl, 0.85 mmol) was added. Thesolution stirred overnight at rt. The mixture was purified via GilsonHPLC to obtain 121 (112 mg, 82%) as a white solid. ^(1H) NMR (300 MHz,CD₃OD): d 8.39 (d, J=9.0 Hz, 1H), 8.10 (d, J=9.3 Hz, 2H), 7.79 (m, 2H),7.63 (s, 1H), 7.58 (s, 1H), 7.39 (d, J=8.8 Hz, 1H), 5.87 (m, 2H), 5.30(d; J=9.6 Hz, 1H), 5.18 (d, J=9.0 Hz, 1H) 4.78 (m, 3H), 4.20 (s, 1H),4.05 (s, 3H), 3.30 (m, 1H), 3.20 (d, 1H), 3.18 (m, 3H), 2.87 (s, 3H),2.80 (m, 1H), 2.50 (m, 1H), 2.10 (m, 1H), 1.62 (m, 2H) 1.40 (m, 2H) 1.05(s, 9H). ³¹P (121.4 MHz, CD₃OD):

37.043.

Example 122 Preparation of Compound 122

Compound 94 (200 mg, 0.27 mmol) was dissolved in ACN (5 mL) followed bythe addition of TEA (1 mL) and the solution was heated at 70° C. for 10min. To the heated solution, isobutylchloro-methylcarbonate was addedand the reaction mixture was further heated for 5 h. The reactionmixture was then cooled to rt and the solvents were removed underreduced pressure. The crude product was purified by reverse phase prepHPLC followed by lyophilization to afford 102 mg of phosphinate prodrugin 49% yield.

¹H NMR (300 MHz, CD₃OD): d 8.01-7.95 (m, 3H), 7.57-7.48 (m, 3H), 7.39(s, 1H), 7.26 (m, 1H), 7.09 (m, 1H), 6.78-6.70 (m, 1H), 5.98-5.55 (m,4H), 5.35-5.10 (m, 2H), 4.74-4.64 (s, 1H), 4.55-4.52 (m, 2H), 4.29-4.26(m, 1H), 4.07-4.03 (m, 1H), 3.95-3.81 (m, 4H), 3.34-3.³¹ (m, 1H),2.69-2.63 (m, 1H), 2.38-2.13 (m, 2H), 1.99-1.33 (m, 12H), 1.³¹-0.80 (m,18H). ³¹P (121.4 MHz, CD₃OD): 53.15 (s, ³¹P); LC/MS: M+1=863.

Example 123 Preparation of Compound 123

Step 1. To dipeptide (1.0 g, 2.70 mmol) dissolved in 20 mL of THF wasadded DSC (1.38 g, 5.40 mmol) followed by NaH (60% dispersion in mineraloil) (270 mg, 6.75 mmol). The reaction was refluxed for 6 hs, quenchedby adding 120 mL of 1M solution of HCl in water, extracted by EtOAc anddried using anhydrous magnesium sulfate. The organic phase wasconcentrated under vacuo, dissolved in 6 mL of DCM and added to amicrowave flask. To the solution was added2-piperidin-1-yl-5-trifluoromethyl-phenylamine (1.98 mg, 8.10 mmol). Themicrowave flask was sealed and put on the microwave apparatus. Thereaction was heated to 65° C. for 1 hour. The reaction was purified bysilica gel chromatography using SiO₂ (eluted with 0% to 100%EtOAc/hexane) to give crude as a yellow solid (1.0 g, 58%). LC/MS=641(M⁺+1)

Step 2. To crude (100 mg, 0.156 mm) dissolved in 1.5 mL of pyridine wasadded NaI (467 mg, 3.12 mm). The reaction was heated to 115° C. for 6hours. After cooled back to rt, pyridine was removed under high vacuo.The crude was dissolved in 2 mL of H₂O and washed by diethyl ether (2×5mL) and was adjusted to pH=2 by adding 3 M HCl solution in water. Thecrude acid was isolated by extracting with EtOAc (2×30 mL) and used fornext step without further purification. To the crude acid was added(1-Amino-2-vinyl-cyclopropyl)-methyl-phosphinic acid (example 1) (50 mg,0.³¹2 mm), HATU (148 mg, 0.390 mm), NMM (79 mg, 0.78 mm) and DMF (5 mL).The reaction was stirring at rt for 12 hours. The reaction solution wasfiltrated and purified by reverse phase HPLC (eluted with 10% to 75%H₂O/CH₃CN) to give 123 as a white solid (45 mg, 37%). ¹H NMR (300 MHz,CD₃CN): d 7.39 (m, 3H), 6.11 (br, 1H), 5.85 (m, 1H), 5.41 (bs, 2H), 5.21(m, 1H), 5.03 (m, 1H), 4.90-4.40 (m, 6H), 4.33 (m, 1H), 3.90 (m, 1H),2.95-2.80 (m, 6H), 2.45-2.35 (m, 2H), 2.17-2.07 (m, 1H), 1.85-1.33 (m,17H), 1.02 (s, 9H). ³¹P NMR (121.4 MHz, CD₃CN):

51.297. LC/MS: 770 (M⁺+1).

Example 124 Preparation of Compound 124

Step 1. The 5-Hydroxy-1,3-dl Hydro-isoindole-2-carboxylic acidtert-butyl ester (2.00 g, 0.85 mmol) and 2-chloro-N,N-dimethylethylaminehydrochloride (1.47 g, 1.02 mmol) were dissolved in 20 mL of CH₃CN.Cesium carbonate (6.92 g, 2.12 mmol) was added and the solution washeated to 65° C. for 18 hours. The mixture was cooled to rt and thesolid was filtered off. The filtrate was concentrated and the residuewas dissolved in 15% MeOH/CH₂Cl₂, washed with H₂O (×2), dried withNa₂SO₄, filtered, and concentrated. The crude product was purified usingsilica gel column chromatography CH_(Z) Cl₂/MeOH to give amine (2.50 g,19%) as a brown waxy solid. ^(1H) NMR (300 MHz, CDCl₃): d 7.16 (m, 1H),6.85 (m, 2H), 5.30 (s, 1H), 4.61 (t, 4H), 4.08 (m, 2H), 2.77 (m, 2H),2.35 (s, 6H), 1.51 (s, 9H).

Step 2. The amine (480 mg, 1.57 mmol) was treated with 5 mL of 4 NHCl/1,4-dioxane and 2 mL of CH₂Cl₂. The reaction mixture was stirredovernight at rt. The solution was concentrated and co-evaporated withtoluene (×2), CHCl₃ and dried under vacuum to give diamine (416 mg, 95%)as a dark solid.

Step 3. Dipeptide,1-(2-Cyclopentyloxycarbonylamino-3,3-dimethyl-butyryl)-4-hydroxy-pyrrolidine-2-carboxylicacid methyl ester (200 mg, 0.54 mmol) was dissolved in 2 mL of CH₂Cl₂and CDI (109 mg, 0.67 mmol) was added. The mixture was stirred at rt for5 hours. A mixture of triethylamine (0.24 mL, 1.72 mmol) and diamine(377 mg, 1.35 mmol) in 1 mL of CH₂Cl₂ was added to the reaction. Thereaction mixture was stirred at rt overnight. The solution wasconcentrated and the product was partitioned between H₂O and 15%MeOH/CH₂Cl₂ (×3). The organic layer was concentrated and purified usinga Gilson HPLC to give ester (277 mg, 85%). ^(1H) NMR (300 MHz, CDCl₃): d7.16 (d, J=8.3 Hz, 1H), 7.08 (d, J=8 Hz, 1H), 6.85 (m, 2H), 6.73 (s,1H), 5.38 (s, 1H), 5.21 (d, J=9.5 Hz, 1H), 4.72 (t, 2H), 4.66 (m, 2H),4.25 (m, 2H), 4.05 (m, 2H), 3.79 (s, 3H), 2.79 (m, 2H), 2.54 (m, 1H),2.47 (s, 6H), 2.22 (m, 1H), 1.61 (m, 1H), 1.55 (m, 2H), 1.45 (m, 2H),1.04 (s, 9H).

Step 4. Ester (275 mg, 0.46 mmol) was dissolved in 4 mL of H₂O/CH₃CN(1/1) and NaOH (183 mg, 4.60 mmol) was added. The reaction mixture wasstirred for 1 hour at rt. The CH₃CN was removed in vacuo and the aqueouslayer was acidified using 1 N HCl. The product was extracted with 15%MeOH/CH₂Cl₂ (×3), dried over Na₂SO₄, filtered and concentrated. Thecrude product (201 mg, 75%) was coupled to phosphinic acid (example 1)to give 124 (52 mg, 21%) as a white foam. ^(1H) NMR (300 MHz, CD₃OD): d7.29 (d, J=8.6 Hz, 1H), 7.19 (d, J=8.2 Hz, 1H), 7.01 (m, 2H), 5.96 (m,1H), 5.³¹ (s, 1H), 5.12 (d, J=10.7 Hz, 1H), 4.68 (t, 2H), 4.50 (t, 1H),4.38 (s, 1H), 4.14 (s, 1H), 3.84 (d, 1H), 3.62 (m, 2H), 3.³¹ (s, 3H),2.98 (m, 6H), 2.44 (m, 1H), 2.21 (m, 1H), 2.18 (m, 1H), 1.61 (m, 1H),1.55 (m, 2H), 1.45 (m, 2H), 1.04 (s, 9H); ³¹P (121.4 MHz, CD₃OD): d47.57.

Example 125 Preparation of Compound 125

Phosphinic acid (10 mg, 1.3 μmol) was dissolved in H₂O (0.2 mL) andtreated with 0.1 N NaOH to adjust pH=11. The mixture was lyophilized,dissolved in NMP (0.3 mL), and heated to 60° C. TEA (7 μL) andisobutylchloromethylcarbonate (19 mg, 0.013 mmol) were added. Thereaction mixture was stirred at 60° C. for 1 h. The reaction mixture wascooled to r.t. and purified by HPLC to give 4.5 mg of 125 in 39% yield.¹H NMR (300 MHz, CD₃OD): d 8.30 (d; J=9.3 Hz, 1H), 8.20 (s, 1H), 7.80(m, 2H), 7.35 (d, J=9.0 Hz, 1H), 6.05-5.60 (m, 4H), 5.40 (m, 1H), 5.20(m, 1H), 4.65 (m, 2H), 4.45 (broad, s, 1H), 4.20-4.00 (m, 7H), 2.80 (m,1H), 2.45 (m, 1H), 2.20 (m, 1H), 1.80-1.45 (m, 10H), 1.40-1.22 (m, 14H),1.00 (m, 10H); ³¹P (121.4 MHz, CD₃OD): d 57.17, 52.94; LC/MS: 913.

Example 126 Preparation of Compound 126

Following experimental procedures similar to those described for thepreparation of compound 125, 18.4 mg of compound 126 was prepared. ^(1H)NMR (300 MHz, CD₃OD): d 7.91-8.15 (m, 5H), 7.³¹-7.66 (m, 7H), 7.23 (s,1H), 7.07 (d, 1H, J=8.7 Hz), 5.79-6.10 (m, 3H), 5.45-5.56 (br m, 1H),5.34 and 5.28 (two d, 1H, J=˜11 Hz), 5.17 and 5.08 (two d, 1H, J=˜11Hz), 4.73 (br m, 1H), 4.46-4.57 (br m, 2H), 4.26 (s, 1H), 3.98-4.08 (m,1H), 3.95 and 3.91 (two s, 3H), 2.56-2.70 (m, 1H), 2.23-2.38 (m, 1H),2.09-2.23 (m, 1H), 1.37-1.83 (m, 13H), 1.05 and 1.02 (two s, 9H). ³¹P(121.4 MHz, CD₃OD): d 57.517, 53.031. LC/MS=867 (M⁺+1)

Example 127 Preparation of Compound 127

Compound 94 (200 mg, 0.27 mmol) was dissolved in ACN (5 mL) followed bythe addition of TEA (1 mL) and the solution was heated at 70° C. for 10min. To the heated solution, isobutylchloromethylcarbonate was added andthe reaction mixture was further heated for 5 h. The reaction mixturewas then cooled to rt and the solvents were removed under reducedpressure. The crude product was purified by reverse phase prep HPLCfollowed by lyophilization to afford 102 mg of phosphinate prodrug in49% yield. ¹H NMR (300 MHz, CDCl₃): d 8.06-8.01 (m, 3H), 7.56-7.48 (m,4H), 7.21-7.01 (m, 3H), 6.14-6.08 (m, 1H), 5.87-4.98 (m, 6H), 4.59-3.97(s, 8H), 3.34-3.³¹ (m, 1H), 2.58-2.52 (m, 2H), 2.20-1.96 (m, 1H),1.70-1.04 (m, 26H). ³¹P (121.4 MHz, CDCl₃): 51.12 (s, ³¹P); LC/MS:M+1=835, M+Na=857.

Example 128 Preparation of Compound 128

Compound 94 (100 mg, 0.14 mmol) was dissolved in pyridine (3 mL)followed by the addition of m-Cl phenol (175 mg, 1.37 mmol) and thesolution was heated at 60° C. for 10 min. To the heated solution wasadded dicyclohexylcarbodiimide (169 mgs, 0.82 mmol) and the reactionmixture was further heated for 3 h. The reaction mixture was then cooledto rt and the solvents were removed under reduced pressure. Dilute thereaction mixture with EtOAc and filter the solids. Remove solvent underreduced pressure and purify the crude product on combi-flash EtOAc/Hexto afford 46 mg of phosphinate prodrug in 40% yield. ¹H NMR (300 MHz,CDCl₃): d 8.10-7.99 (m, 2H), 7.57-6.99 (m, 10H), 5.89-5.83 (m, 1H),5.41-4.93 (m, 4H), 4.73-3.96 (m, 5H), 3.15-2.80 (m, 2H), 2.56 (m, 1H),2.05-0.91 (m, 27H). ³¹P (121.4 MHz, CDCl₃): 51.12 (s, ³¹P); LC/MS:M+1=843.

Example 129 Preparation of Compound 129

Acid compound 58 (128 mg, 0.14 mmol) was dissolved in CH₃CN (2.5 mL) andheated to 65° C. for 10 minutes. TEA (0.2 mL, 1.41 mmol) and BOMCl (480mg, 2.82 mmol) were added. The reaction mixture was stirred at 65° C.for 24 h and cooled to rt. The reaction was quenched with H₂O andorganic solvents were evaporated. The aqueous layer was extracted withEtOAc. The aqueous layer was brought to pH=2 and extracted with EtOAc.The combined organic layers were dried with Na₂SO₄, filtered andconcentrated. The crude product was purified by HPLC to give 10 mg of129.

Example 130 Preparation of Compound 130

To a solution of compound 35 (725 mg, 0.831 mmol) in CH₃CN (20 mL) wasadded TEA (1.16 mL, 0.831 mmol) and the solution was heated at 70° C.for 10 min. POC—Cl was then added to the reaction mixture, and heatingwas continued for 5 h. The mixture was concentrated under reducedpressure and purified on reverse phase HPLC to afford 219 mgs of thephosphinate prodrug in 27% yield. ¹H NMR (300 MHz, CD₃OD): d 8.89 (s,1H), 8.30 (m, 1H), 8.18 (s, 1H), 7.76 (s, 1H), 7.35-7.23 (m, 6H),6.03-5.77 (m, 2H), 5.57-5.28 (m, 3H), 5.15-5.01 (m, 2H), 4.86-4.65 (m,2H), 4.45 (s, 1H), 4.22-4.05 (m, 5H), 3.65-3.20 (m, 2H), 2.81-2.74 (m,2H), 2.50-2.44 (m, 2H), 2.18-2.15 (m, 1H), 1.77-1.23 (m, 23H), 1.19-0.97(m, 10H). ³¹P (121.4 MHz, CD₃OD): 48.55; LC/MS: M+1=989.

Example 131 Preparation of Compound 131

Following experimental procedures similar to those described for thepreparation of compound 130, 15 mg of compound 131 was prepared. ^(1H)NMR (300 MHz, CD₃OD): d 8.05 (d, 1H, J=9.6 Hz), 7.48 and 7.46 (two s,1H), 7.43 (s, 1H), 7.36 (d, 1H), 7.04 (dd, 1H, J=9.6 Hz), 5.76-6.06 (m,1H), 5.56-5.76 (m, 2H), 5.48 (br, 1H), 5.26-5.38 (m, 1H), 5.14 (appt t,1H, J=˜12 Hz), 4.78 (br, 1H), 4.46-4.57 (m, 2H), 4.28 (s, 1H), 4.06 (brd, 1H, J=˜11 Hz), 3.95 (s, 3H), 3.88-4.00 (m, 1H), 3.80 and 3.72 (two s,3H), 2.94 (br m, 0.5H), 2.62-2.75 (m, 1.5H), 2.22-2.42 (m, 1H),2.06-2.22 (m, 1H), 1.42-1.84 (m, 10H), 1.32 (d, 6H, J=6.6 Hz), 1.27-1.36(m, 1H), 1.20 (appt t, 1H, J=7.4 Hz), 1.06 and 1.04 (two s, 9H). ³¹P(121.4 MHz, CD₃OD): d 57.608, 53.232. LC/MS=885 (M⁺+1)

Example 132 Preparation of Compound 132

Phosphinic acid (500 mg, 5.73 mmol) and alcohol (1.87 g, 57.3 mmol) weredissolved in DMF (3 mL). PyBop (843 mg, 20.06 mmol), TEA (0.4 mL, 28.65mmol), and DMAP (14 mg, 1.15 mmol) were added. The reaction mixture wasstirred at r.t. for 3 h and concentrated. The product was partitionedbetween brine and CH₂Cl₂ (3×). The organic layer was dried with Na₂SO₄and concentrated. The residue was purified by combi-flash to give 406 mgof intermediate silyl ether in 60% yield. The resulting silyl ether (406mg, 3.44 mmol) was dissolved in THF (3 ML) and 1.0 M TBAF in THF (0.43mL, 4.3 mmol) was added. The reaction mixture was stirred for 1 h andconcentrated. The product was partitioned between H₂O and CH₂Cl₂. Theorganic layer was concentrated and purified by HPLC to give 227 mg of132 in 70% yield. ¹H NMR (300 MHz, CDCl₃): d 8.80 (s, 1H), 8.20 (m, 1H),7.80 (m, 2H), 7.65 (m, 3H), 7.45-7.17 (m, 6H), 5.80-5.65 (m, 2H),5.40-5.05 (m, 4H), 4.65 (m, 2H), 4.40-3.95 (m, 8H), 3.60-3.20 (m, 3H),2.70 (m, 1H), 2.00 (m, 1H), 1.80-1.35 (m, 13H), 1.05-0.95 (m, 16H); ³¹P(121.4 MHz, CDCl₃): d 50.24, 48.92; LC/MS: 943.

Example 133 Preparation of Compound 133

Step 1. 4,5-dimethyl-2-oxo-1,3-dioxole (5 g, 43.82 mmol), NBS (8.19 g,46.01 mmol), and benzoyl peroxide (20 mg) were dissolved in CCl₄ (30 mL)and heated to 80° C. for 1.5 h. The reaction mixture was cooled to r.tand the solid was filtered off. The filtrate was concentrated. Theresidue was purified by silica gel column to give 8.29 g of4-bromomethyl-5-methyl-2-oxo-1,3-dioxole as a yellow oil. TEA (12 mL,86.1 mmol) was added dropwise to a solution of4-bromomethyl-5-methyl-2-oxo-1,3-dioxole (6 g, ³¹.09 mmol) and formicacid (3.36 mL) in CH₃CN (96 mL) while keeping the temperature under 20°C. The mixture was stirred at r.t. for 2 h and concentrated. The productwas partitioned between H₂O and EtOAc (3×). The organic layer was driedwith Na₂SO₄, concentrated and dried in vacuo to give crude formate. Theresulting formate was dissolved in MeOH (40 mL) and 0.5 mL ofconcentrated HCl was added. The mixture was stirred at r.t. for 5 h,concentrated, and co-evaporated with toluene. The crude product waspurified by silica gel column to give 2.8 g of4-hydroxymethyl-5-methyl-2-oxo-1,3-dioxole in 69% yield. ¹H NMR (300MHz, CDCl₃): d 4.40 (s, 2H), 2.60 (broad, s, 1H), 2.20 (s, 3H).

Phosphinic acid (150 mg, 0.17 mmol) and4-hydroxymethyl-5-methyl-2-oxo-1,3-dioxole (112 mg, 0.85 mmol) weredissolved in DMF (1 mL). PyBop (179 mg, 0.34 mmol), TEA (0.07 mL, 0.51mmol), and DMAP (7 mg) was added. The mixture was stirred at r.t.overnight. The product was partitioned between aqueous NaHCO₃ and EtOAc(3×). The organic layer was washed with NH₄Cl and brine, dried withNa₂SO₄, and concentrated. The crude product was purified by HPLCfollowed by silica gel column to give 40 mg of (133) in 24% yield. ¹HNMR (300 MHz, CDCl₃): d 8.00 (d, J=9.3 Hz, 1H), 7.5 (s, 1H), 7.40 (m,2H), 7.30-7.20 (m, 5H), 7.00 (d, J=8.7 Hz, 1H), 5.95-5.80 (m, 2H),5.40-5.10 (m, 5H), 5.00 (broad, s, 1H), 4.70-4.40 (m, 5H), 4.00 (s, 5H),3.70 (m, 1H), 3.30 (m, 2H), 2.80 (m, 1H), 2.60 (m, 1H), 2.00-1.35 (m,16H), 1.05 (m, 12H); ³¹P (121.4 MHz, CDCl₃): d 50.81, 47.39; LC/MS:M+1=985

Example 134 Preparation of Compound 134

Step 1. The 2-tert-butoxycarbonylamino-pentanedioic acid 1-benzyl ester(4.06 g, 12 mmol) and TEA (5 mL, 35.87 mmol) were dissolved in THF (60mL) and cooled to 0° C. Ethylchloroformate (3.4 mL, 35.7 mmol) was addeddropwise. The mixture was stirred at 0° C. for 5 minutes and warmed tor.t. for 1 h. NaBH₄ (1.88 g, 49.7 mmol) was added followed by additionof 1 drop of H₂O. The reaction mixture was stirred at r.t. overnight. 4N HCl was added at 0° C. and extracted with EtOAc (100 mL). The aqueouslayer was washed with H₂O (100 mL), NaOH (2×100 mL), H₂O (100 mL) andbrine (100 mL). The organic layer was dried with Na₂SO₄, filtered, andconcentrated. The crude product was purified by combi-flash to give 2.89g of 2-tert-butoxycarbonylamino-5-hydroxy-pentanoic acid benzyl ester in75% yield.

To a solution of alcohol 2-tert-butoxycarbonylamino-5-hydroxy-pentanoicacid benzyl ester (1.64 g, 5.07 mmol) in ether (15 mL) was added Ag₂O(4.08 g, 17.61 mmol) and allyl bromide (2.6 mL, 29.87 mmol). Thereaction mixture was stirred at r.t. overnight. The reaction mixture wasfiltered through celite and concentrated. The crude product was purifiedby combi-flash to give 940 mg of5-allyloxy-2-tert-butoxycarbonylamino-pentanoic acid benzyl ester.

Step 2. To a solution of 5-allyloxy-2-tert-butoxycarbonylamino-pentanoicacid benzyl ester (5.95 g, 16.38 mmol) in CH₂Cl₂ (100 mL) was added 4 NHCl in 1,4-dioxane (100 mL, 400 mmol). The reaction mixture was stirredat r.t. for 2 h, concentrated, and dried under vacuo to give amine HClsalt. The resulting amine HCl salt was dissolved in THF (150 mL) and H₂O(25 mL). TEA (7 mL, 50.2 mmol) and carbonic acid cyclopentyl ester2,5-dioxo-pyrrolidin-1-yl ester (3.92 g, 17.25 mmol) were added. Thereaction mixture was stirred at r.t. for 1 h. H₂O (200 mL) was added andthe organic solvent was reduced on rotavap. The remaining mixture wasextracted with EtOAc (3×150 mL). The combined organic layer was washedwith 1 N HCl, H₂O and brine, dried with Na₄SO₄, concentrated, and driedto give 6.41 g of ester as crude product.

Ester (6.41 g, 17.07 mmol) was dissolved in THF (65 mL)/H₂O (75 mL), andLiOH (1.63 g, 38.85 mmol) was added. The reaction mixture was stirred atr. t. overnight and diluted with EtOAc. The reaction mixture wasacidified to pH=2 with 1 N HCl and separated. The aqueous layer wasextracted with EtOAc (3×150 mL). The combined organic layers were washedwith brine, dried with Na₂SO₄, concentrated and dried under vacuum togive 4.87 g of 5-allyloxy-2-cyclopentyloxycarbonylamino-pentanoic acid.

Step 3. To a solution of intermediate XII (2.25 g, 3.07 mmol) in CH₂Cl₂(20 mL) was added 4 N HCl in 1,4-dioxane (20 mL, 80 mmol). The reactionmixture was stirred at r.t. for 1 h, concentrated, and dried under vacuoto give amine HCl salt. The resulting amine HCl salt and acid (1.05 g,3.67 mmol) were dissolved in DMF (30 mL). HATU (2.36 g, 6.20 mmol) andNMM (1.56 g, 15.46 mmol) were added and the mixture was stirred at r.t.for 2 h. The reaction was diluted with EtOAc and washed with 20% LiCl(2×100 mL). The organic layer was washed with aqueous NH₄Cl (200 mL),dried with Na₂SO₄, and concentrated. The crude product was purified bycombi-flash to give 2.5 g of diene in 94% yield.

Step 4. Diene (2.59 g, 2.87 mmol) was dissolved in CH₂Cl₂ (300 mL) anddegassed with N₂ for 20 minutes. Grubb's G1 (664.5 mg, 0.73 mmol) wasadded and degassed for an additional 20 minutes. The reaction mixturewas heated to 45° C. overnight and cooled to rt.Tris(hydroxymethyl)phosphine (5.03 g, 40.54 mmol) was added followed byaddition of TEA (11.2 mL, 80.35 mmol) and H₂O (47 mL). The reactionmixture was heated to 45° C. for 4 h and then r.t. overnight. The twolayers were separated. The organic layer was washed with 0.5 N HCl,brine, dried with Na₂SO₄, and concentrated. The crude product waspurified by combi-flash to give 1.03 g of macrocyclic phosphinate.

Step 5. A solution of phosphinate (1.00 g, 1.15 mmol) and8-chloro-2-(2-isopropylamino-thiazol-4-yl)-7-methoxy-quinolin-4-ol (469mg) in NMP (12 mL) was treated with Cs₂CO₃ (1.20 g). The reactionmixture was heated to 70° C. overnight and then cooled to rt. Thereaction was diluted with 5% LiCl (150 mL) and extracted with EtOAc(3×150 mL). The combined organic layers were washed with brine, driedwith Na₂SO₄, and concentrated. The crude product was purified bycombi-flash to give 964.3 mg of desired product.

To a solution of product obtained above (964.3 mg, 0.98 mmol) in DME (10mL)/H₂O (1 mL) was added p-tosylhydrazide (1.37 g, 7.36 mmol) and NaOAc(1.22 g, 14.8 mmol). The reaction mixture was heated to 95° C. for 2 hand cooled to rt. The mixture was diluted with EtOAc (125 mL) and washedwith saturated NaHCO₃ (2×50 mL). The aqueous layer was extracted withEtOAc (25 mL). The combined organic layer was washed with brine (25 mL),dried with Na₂SO₄, and concentrated. The crude product was purified bycombi-flash to give 437.6 mg of 134. ¹H NMR (300 MHz, CD₃OD): d 8.36 (d,J=8.9 Hz, 1H), 8.19 (s, 1H), 7.90 (s, 1H), 7.64 (d, J=9.7 Hz, 1H), 7.2(m, 1H), 6.86 (m, 2H), 5.9 (bs, 1H), 4.77 (m, 1H), 4.63 (m, 1H), 4.40(m, 1H), 4.3-3.95 (m, 8H), 3.60 (bs, 3H), 3.55-3.35 (m, 5H), 2.81 (m,1H), 2.68 (m, 1H) 2.15-1.1 (m, 25H). LC/MS=959.29 (M⁺+1 LC/MS=959.29(M⁺+1).

Example 135 Preparation of Compound 135

The fully protected phosphinate (synthesized as described in example 58with Boc protection group) was treated with HCl to remove the Bocprotection group. The resulting amine was used to prepare compound135-141.

To a solution of this amine (390 mg, 0.47 mmol) in EtOAc (50 mL) wasadded saturated NaHCO₃ (60 mL) and stirred vigorously. Cyclopentylacetylchloride (76 mg, 0.52 mmol) in EtOAc (1 mL) was added and stirredfor 15 minutes. The two layers were separated. The organic layer waswashed with brine and concentrated. The dried residue was dissolved inCH₃CN (5 mL) and cooled to 0° C. Iodotrimethylsilane (0.60 mL, 2.37mmol) was added. The reaction mixture was warmed to rt, stirred for 0.5h, and cooled to 0° C. 2,6-lutidine (1.5 mL, 4.73 mmol) was addedfollowed by addition of MeOH (5 mL). The mixture was concentrated invacuo. The residue was purified by HPLC to give 337.2 mg of compound135. ¹H NMR (300 MHz, CD₃OD): d 8.25 (d, J=9.0 Hz, 1H), 8.15 (s, 1H),7.76 (s, 1H), 7.73 (d, J=2.1 Hz, 1H), 7.32 (dd, J=2.4, 9.0 Hz, 1H), 7.23(t, J=7.8 Hz, 1H), 6.90 (t, J=7.8 Hz, 2H), 5.99 (m, 1H), 5.78 (brs, 1H),5.30 (d, J=15.3 Hz, 1H), 5.12 (d, J=11.7 Hz, 1H), 4.67 (m, 2H), 4.50 (m,1H), 4.14 (m, 2H), 4.05 (s, 3H), 3.40 (m, 2H), 2.75 (m, 1H), 2.63 (m,1H), 2.0-2.4 (m, 3H), 1.91 (m, 1H), 1.4-1.7 (m, 8H), 1.34 (d, J=6.3 Hz,6H), 0.95-1.15 (brs, 1H); ³¹P (121.4 MHz, CD₃OD):

42.091; LC/MS=907 (M⁺+1).

Example 136 Preparation of Compound 136

Following experimental procedures similar to those described for thepreparation of compound 135, compound 136 was prepared. ¹H NMR (300 MHz,CD₃OD): d 8.25 (d, J=9.0 Hz, 1H), 8.17 (s, 1H), 7.76 (s, 1H), 7.75 (d,J=2.1 Hz, 1H), 7.33 (dd, J=2.1, 9.0 Hz, 1H), 7.26 (t, J=7.8 Hz, 1H),6.93 (t, J=7.8 Hz, 2H), 5.97 (m, 1H), 5.80 (brs, 1H), 5.³¹ (d, J=15.9Hz, 1H), 5.12 (d, J=12.0 Hz, 1H), 4.66 (m, 2H), 4.49 (t, 1H), 4.14 (m,2H), 4.06 (s, 3H), 3.41 (d, 2H), 2.77 (m, 1H), 2.56 (m, 1H), 2.22 (m,1H), 2.06 (m, 2H), 1.62 (m, 1H), 1.47 (m, 1H), 1.2-1.4 (m, 8H), 1.07 (s,9H), 0.80 (t, J=7.4 Hz, 3H); ³¹P (121.4 MHz, CD₃OD):

41.043.

Example 137 Preparation of Compound 137

Following experimental procedures similar to those described for thepreparation of compound 135, compound 137 was prepared. ¹H NMR (300 MHz,CD₃OD): d 8.26 (d, J=9.0 Hz, 1H), 8.17 (s, 1H), 7.76 (s, 1H), 7.75 (d,J=2.7 Hz, 1H), 7.32 (dd, J=2.7, 9.2 Hz, 1H), 7.26 (t, J=7.8 Hz, 1H),6.93 (t, J=7.8 Hz, 2H), 5.97 (m, 1H), 5.81 (brs, 1H), 5.³¹ (dd, J=1.4,17.0 Hz, 1H), 5.12 (dd, J=1.4, 10.5 Hz, 1H), 4.70 (m, 2H), 4.41 (t, 1H),4.14 (m, 2H), 4.05 (s, 3H), 3.41 (d, 2H), 2.79 (m, 1H), 2.57 (m, 1H),2.33 (m, 2H), 2.19 (m, 1H), 2.04 (m, 2H), 1.64 (m, 1H), 1.46 (m, 1H),1.34 (d, J=6.3 Hz, 6H), 1.06 (s, 9H); ³¹P (121.4 MHz, CD₃OD):

41.952.

Example 138 Preparation of Compound 138

Following experimental procedures similar to those described for thepreparation of compound 135, compound 138 was prepared. ¹H NMR (300 MHz,CD₃OD): d 8.25 (d, J=9.3 Hz, 1H), 8.18 (s, 1H), 7.76 (s, 2H), 7.33 (dd,J=2.4, 9.0 Hz, 1H), 7.26 (t, J=7.8 Hz, 1H), 6.93 (t, J=7.8 Hz, 2H), 5.97(m, 1H), 5.79 (brs, 1H), 5.32 (d, J=15.9 Hz, 1H), 5.13 (d, J=12.0 Hz,1H), 4.67 (m, 2H), 4.48 (t, 1H), 4.14 (m, 2H), 4.05 (s, 3H), 3.42 (d,2H), 2.78 (m, 1H), 2.57 (m, 1H), 2.20 (m, 1H), 1.98 (d, J=12.6 Hz, 1H),1.90 (d, J=12.6 Hz, 1H), 1.64 (m, 1H), 1.48 (m, 1H), 1.34 (d, J=9.6 Hz,6H), 1.06 (s, 9H, 0.80 (s, 9H); ³¹P (121.4 MHz, CD₃OD):

41.077; LC/MS=895 (M⁺+1).

Example 139 Preparation of Compound 139

Following experimental procedures similar to those described for thepreparation of compound 135, compound 139 was prepared. ¹H NMR (300 MHz,CD₃OD): d 8.27 (d, J=9.6 Hz, 1H), 8.16 (s, 1H), 7.74 (s, 2H), 7.34 (dd,J=2.1, 9.3 Hz, 1H), 7.25 (t, J=7.8 Hz, 1H), 6.92 (t, J=7.8 Hz, 2H), 5.96(m, 1H), 5.79 (brs, 1H), 5.30 (d, J=17.1 Hz, 1H), 5.12 (d, J=10.5 Hz,1H), 4.72 (t, 1H), 4.52 (d, 1H), 4.14 (m, 2H), 4.05 (s, 3H), 3.41 (d,2H), 3.00 (t, J=8.3 Hz, 2H), 2.59 (m, 3H), 2.19 (m, 1H), 2.04 (m, 2H),1.63 (m, 1H), 1.47 (m, 1H), 1.34 (d, J=6.3 Hz, 6H), 1.10 (s, 9H); ³¹P(121.4 MHz, CD₃OD):

38.19

40.829.

Example 140 Preparation of Compound 140

Step 1. Ethyl acetimidate hydrochloride (1.23 g, 9.95 mmol) and2,2,2-trifluoroethylamine hydrochloride (1.35 g, 9.95 mmol) weredissolved in CH₂Cl₂ (32 mL)/H₂O (3.2 mL). K₂CO₃ (0.69 g, 4.98 mmol) wasadded and stirred for 30 minutes. The two layers were separated. Theaqueous layer was extracted with CH₂Cl₂ (2×10 mL). The combined organiclayer was dried with Na₂SO₄ and concentrated to give 1.48 g of thedesired amidate as a light yellow liquid in 87% yield.

Step 2. The phosphinate (500 mg, 0.54 mmol) was dissolved in CH₃CN (5mL) and cooled to 0° C. Iodotrimethylsilane (0.77 mL) was added. Thereaction mixture was warmed to rt, stirred for 0.5 h, and cooled to 0°C. 2,6-lutidine (1.30 mL) was added followed by addition of MeOH (5 mL).The mixture was concentrated, co-evaporated with CH₂Cl₂ (2×), and driedin vacuo to give the desired amino phosphinic acid as the 2,6-lutidinesalt.

Step 3. The amino phosphinic acid obtained from step 2 (80 mg, 0.025mmol) was dissolved in DMF (0.45 mL) and 0.1 N phosphate buffer (0.9mL). 2 N NaOH (86 μL) was added to adjust pH to 9. A solution of theamidate obtained from step 1 (150 mg, 0.89 mmol) in DMF (0.1 mL) wasadded and stirred for 18 h. The reaction mixture was filtered and thefiltrate was purified by HPLC to give 8.8 mg of compound 140. ¹H NMR(300 MHz, CD₃OD): d 8.27 (d, J=9.6 Hz, 1H), 8.16 (s, 1H), 7.74 (s, 2H),7.34 (dd, J=2.1, 9.3 Hz, 1H), 7.25 (t, J=7.8 Hz, 1H), 6.92 (t, J=7.8 Hz,2H), 5.97 (m, 1H), 5.79 (brs, 1H), 5.30 (d, J=17.1 Hz, 1H), 5.12 (d,J=10.5 Hz, 1H), 4.72 (t, 1H), 4.52 (d, 1H), 4.14 (m, 2H), 4.05 (s, 3H),3.41 (d, 2H), 3.³¹ (s, 3H), 3.01 (t, J=8.3 Hz, 2H), 2.80 (m, 1H), 2.59(m, 5H), 2.19 (m, 1H), 1.63 (m, 1H), 1.47 (m, 1H), 1.34 (d, J=6.3 Hz,6H), 1.10 (s, 9H); ³¹P (121.4 MHz, CD₃OD):

40.829.

Example 141 Preparation of Compound 141

Step 1. A mixture of a-ketoester Oxo-phenyl-acetic acid methyl ester(820 mg, 5 mmol) and Deoxo-Fluor (2.43 g, 11 mmol) was heated to 45° C.and stirred under N₂ for 16 h. The mixture was cooled to rt, poured intoice water, and added CH₂Cl₂ (40 mL). The CH₂Cl₂ layer was collected andconcentrated. The crude product was purified by combi-flash to give 536mg of the corresponding difluoro ester as colorless oil. To a solutionof the difluoro ester (536 mg, 2.88 mmol) in toluene (20 mL) at −78° C.was added 1.0 M DIBAL in CH₂Cl₂ and stirred for 2 h at −78° C. Thereaction mixture was poured into ice cold 6 N HCl (100 mL) and extractedwith CH₂Cl₂. The organic layers were filtered through celite,concentrated to a volume of 40 mL, and used for the next step reaction.

Step 2. To a solution of amino phosphinic acid (example 140, step 2) (65mg, 0.02 mmol) in CH₂Cl₂ (1 mL) was added a solution of aldehydeobtained from step 1 in CH₂Cl₂/toluene (1 mL). TFA (50 μL) andNaBH(OAc)₃ (21 mg) were added and stirred for 16 h. AdditionalNaBH(OAc)₃ (63 mg) and the solution of aldehyde in CH₂Cl₂/toluene (2 mL)were added. The reaction mixture was stirred for 24 h. The reaction wasdiluted with CH₂Cl₂ (30 mL) and washed with saturated NaHCO₃. Theorganic layers were washed with 0.1 N HCl and concentrated. The residuewas dissolved in CH₂Cl₂, filtered through Acrodisk, and concentrated.The crude product was purified by HPLC to give 28.2 mg of (141). ¹H NMR(300 MHz, CD₃OD): d 8.15 (s, 1H), 8.10 (d, J=9.3 Hz, 1H), 7.76 (s, 1H),7.56 (s, 1H), 7.44 (d, J=7.2 Hz, 1H), 7.33 (t, J=8.1 Hz, 2H), 7.26 (t,J=7.8 Hz, 1H), 7.05-7.20 (m, 3H), 6.92 (t, J=7.8 Hz, 2H), 5.97 (m, 1H),5.84 (brs, 1H), 5.³¹ (d, J=16.8 Hz, 1H), 5.12 (d, J=10.5 Hz, 1H), 4.85(m, 2H), 4.43 (d, 1H), 4.13 (m, 2H), 3.98 (s, 3H), 3.41 (d, 2H), 3.22(m, 2H), 2.85 (m, 1H), 2.60 (m, 1H), 2.21 (m, 1H), 1.65 (m, 1H), 1.50(m, 1H), 1.34 (d, J=6.3 Hz, 6H), 1.09 (s, 9H); ³¹P (121.4 MHz, CD₃OD):

40.804.

Example 142 Preparation of Compound 142

To a solution of 92 (600 mg, 0.61 mmol) in DME (9.1 mL) and H₂O (1.02mL) was added p-tosylhydrazide (856 mg, 4.57 mmol) and NaOAc (749 mg,9.14 mmol). The reaction mixture was heated to 95° C. for 3 h and cooledto room temperature. The mixture was concentrated, dissolved in CH₂Cl₂,washed with H₂O, and then slightly acidic H₂O. The organic layer wasdried over Na₂SO₄ and concentrated. The crude product was purified byreverse phase combi-flash followed by HPLC to give acid 142 (366 mg,61%) as a yellow solid. ¹H NMR (300 MHz, CD₃OD)

8.31 (s, 1H), 8.30 (d, J=9 Hz, 1H), 7.87 (s, 1H), 7.67. (d, J=9.3 Hz,1H), 7.28 (m, 1H), 6.96 (t, J=7.8 Hz, 2H), 5.84 (s, 1H), 4.95 (dd, J=6,10.2 Hz, 1H), 4.76 (d, J=11.4 Hz, 1H), 4.71 (d, J=8.4 Hz, 1H), 4.35 (d,J=10.5 Hz, 1H), 4.20 (m, 2H), 4.17 (s, 3H), 4.04 (quint., J=6.6 Hz, 1H),3.49 (t, J=15 Hz, 1H), 3.33, (t, J=15 Hz, 1H), 2.88 (dd, J=7.5, 14.7 Hz,1H), 2.56-2.74 (brm, 2H), 2.29 (m, 1H), 1.92 (m, 2H), 1.40-1.79 (brm,10H), 1.37 (dd, J=1.8, 6.6 Hz, 6H), 1.31 (m, 4H), 1.23 (d, J=6 Hz, 3H),1.00 (m, 1H).

³¹P NMR (121.4 MHz, CD₃OD) d 39.8. LC/MS=987.1 (M⁺+1)

Example 143 Preparation of Compound 143

Step 1. A mixture of the intermediate VIII (1.96 g, 5.36 mmol),benzyl-(1-benzyloxycarbonylamino-2-vinyl-cyclopropyl)-phosphinic acidethyl ester (730 mg, 2.75 mmol), sodium 2-ethylhexanoic acid (300 mg),toluene (5 mL) and water (10 mL) was stirred at 80° C. for 16 h. Theresulting reaction mixture was partitioned between ethyl acetate and 5%aqueous sodium bicarbonate. The organic layer was washed sequentiallywith ice-cold 0.5 N NaOH, 1 N HCl and brine, and then concentrated,affording the desired product (1.15 g, 66%) as an oil.

Step 2. To a solution of the alcohol obtained from step 1 (1.15 g, 1.82mmol) and DABCO (410 mg, 3.66 mmol) in toluene (2 mL) was added asolution of 4-bromobenzenesulfonyl chloride (840 mg, 3.38 mmol) intoluene (2 mL) and stirred at rt for 1 h. The reaction mixture wasdiluted with ethyl acetate, washed sequentially with ice-cold 0.5 NNaOH, 1N HCl and brine and then concentrated, affording the brosylatedproduct (1.38 g, 89%).

Step 3. A solution of the intermediate obtained from step 2 (589 mg,0.69 mmol) in CH₂Cl₂ (60 mL) was purged with nitrogen for 5 min. Aruthenium catalyst (G 1, 110 mg, 0.14 mmol) was added and the mixturewas stirred at 50° C. for 16 h. After cooling,tri(hydroxymethyl)phosphine (860 mg, 6.9 mmol), triethylamine (0.96 mL,6.9 mmol) and water (20 mL) were added and the mixture was vigorouslystirred for 4 h. The organic layer was taken, washed with 1N HCl andthen brine, and concentrated. The residue was purified by Combiflash,affording 328 mg of the desired macrocycle as oil.

Step 4. A mixture of the brosylated compound obtained from step 3 (348mg, 0.42 mmol),2-(2-isopropylamino-thiazol-4-yl)-7-methoxy-quinolin-4-ol (133 mg, 0.42mmol) and cesium carbonate (210 mg, 0.63 mmol) in NMP (3 mL) was stirredat 60° C. for 3 h. Acetic acid (0.1 mL) was added and the mixture wasdiluted with water (0.5 mL) and DMF (2 mL). The crude solution wassubjected to HPLC purification, affording the desired product (132 mg,35%).

Step 5. To a suspension of the compound obtained from step 4 (170 mg,0.19 mmol) in CH₃CN (10 mL) was added bromotrimethylsilane (0.2 mL). Thereaction mixture was stirred at rt for 3 h and then heated to 50° C. for1 h. The reaction was cooled to r.t. and MeOH (1 mL) was added. Themixture was concentrated and the residue was dissolved in DMF (5 mL).The crude product was purified by HPLC and the desired fractionscontaining the desired amino phosphinic acid were combined, andconcentrated to a volume of 20 mL, which was used for the next reaction.

Step 6. The compound obtained from step 5 was in acidic water.Triethylamine (1 mL) was added to bring the solution to pH=9.Cyclopentyl chloroformate (0.1 mL) was added dropwise while stirringuntil the starting material was completely converted to product. Themixture was concentrated to a volume of 10 mL. The product waspartitioned between EtOAc and saturated NH₄Cl. The aqueous layer wasextracted with EtOAc. The combined organic layers were concentrated. Theresidue was purified by HPLC to give 16 mg of compound 143 as a yellowfluffy powder. ¹H NMR (300 MHz, CD₃OD):

8.33 (d, J=9.0 Hz, 1H), 8.18 (s, 1H), 7.74 (s, 1H), 7.73 (s, 1H),7.13-7.33 (m, 6H), 5.83 (brs, 1-H), 5.70 (dd, J=8.6, 18.3 Hz, 1H), 5.28(t, J=9.6 Hz, 1H), 4.73 (t, J=8.7 Hz, 1H), 4.40 (brs, 1H), 4.03-4.22 (m,3H), 4.02 (s, 3H), 3.39 (t, J=15.9 Hz, 1H), 3.18 (t, J=15.9 Hz, 1H),2.80 (m, 1H), 2.61 (m, 1H), 2.26 (m, 1H), 1.82 (m, 3H), 1.2-1.7 (m,23H); ³¹P (121.4 MHz, CD₃OD):

42.259; LC/MS=885 (M⁺+1).

Example 144 Preparation of Compound 144

Step 1. To a solution of the phosphinate (step 4, example 143, 132 mg,0.15 mmol) and 2,4,6-triisopropylbenzenesulfonylhydrazide (440 mg) inTHF (10 mL) was added TEA (0.2 mL). The reaction mixture was heated to55° C. for 4 h. An additional hydrazide (440 mg) and TEA (0.2 mL) wereadded and stirred for 16 h at 55° C. More hydrazide (400 mg) and TEA(0.2 mL) were added and stirred for 4 h. The reaction mixture wasconcentrated and the residue was chromatographed affording a fractioncontaining a mixture of the starting material, the desired product, andthe benzenesulfinic acid byproduct. After removal of the solvents, thematerial was re-dissolved in THF. The hydrazide (440 mg) and TEA (0.4mL) were sequentially added. The reaction mixture was stirred at 55° C.for 16 h. To drive the reaction to completion, additional hydrazide (660mg) and TEA (0.4 mL) were added, and stirred for 6 h at 55° C. Thereaction mixture was concentrated and the residue was chromatographedaffording a fraction containing the desired product. After removal ofthe solvents, the residue was partitioned between CH₂Cl₂ (100 mL) andice-cold 0.5 N NaOH (20 mL). The CH₂Cl₂ layer was washed with 0.5 N NaOH(20 mL) and brine (30 mL) and concentrated. 93 mg of the desiredsaturated macrocycle was obtained as a yellow solid.

Step 2. To a solution of the saturated macrocycle obtained from step 1(93 mg, 0.1 mmol) in CH₂Cl₂ (10 mL) was added bromotrimethylsilane (0.2mL) and stirred at r.t for 2 h. 2,6-Lutidine (0.1 mL) was added andstirred at r.t for 16 h. LCMS showed the desired product with tert-Bocprotected phosphinic acid as a minor product. Additionalbromotrimethylsilane (0.1 mL) was added. The mixture was heated toreflux for 1 h. MeOH (3 mL) was added and concentrated. The crudeproduct was dissolved in DMF (5 mL) and H₂O (0.5 mL) and purified byHPLC to give 27 mg of the desired amino phosphinic acid as a fluffysolid.

Step 3. To a solution of the amino phosphinic acid obtained from step 2and triethylamine (0.2 mL) in CH₃CN (10 mL) and H₂O (1 mL) was addedcyclopentyl chloroformate (50 μL). The reaction mixture was stirred atr.t. for 0.5 h. Additional cyclopentyl chloroformate (50 μL) was addedand stirred for 1 h. After removal of the solvents, the residue wasdissolved in DMF (1.8 mL) and H₂O (0.2 mL) and purified by HPLC to give12 mg of compound 144.

¹H NMR (300 MHz, CD₃OD):

8.28 (d, J=9.0 Hz, 1H), 8.18 (s, 1H), 7.75 (s, 1H), 7.74 (s, 1H),7.13-7.33 (m, 6H), 5.82 (brs, 1H), 4.85 (d, J=12.3 Hz, 1H), 4.68 (t,J=8.7 Hz, 1H), 4.24 (brs, 1H), 4.04-4.23 (m, 3H), 4.03 (s, 3H), 3.34 (t,J=15.9 Hz, 1H), 3.22 (t, J=15.9 Hz, 1H), 2.78 (m, 1H), 2.49 (m, 1H),1.1-2.0 (m, 27H); ³¹P (121.4 MHz, CD₃OD):

44.791; LC/MS=887 (M⁺+1).

Example 145 Preparation of Compound 145

To a yellow solution of VIII (1.67 g, 4.56 mmol) and amine(1-amino-2-vinyl-cyclopropyl)-(2-fluoro-benzyl)-phosphinic acid ethylester (972 mg, 3.5 mmol) in toluene (10 mL) was added a solution ofsodium 2-ethyl hexanoate (871 mg, 5.25 mmol) in H₂O (30 mL). Thereaction mixture was heated to 80° C. overnight and cooled to rt. Thereaction mixture was diluted with EtOAc, washed with saturated NaHCO₃,0.5 N HCl, brine, dried with Na₂SO₄, and concentrated. The crude productwas purified by combi-flash to give 1.47 g of alcohol in 65% yield. ¹HNMR (300 MHz, CDCl₃):

7.6

(

1H) 7.43-7.34 (m, 2H), 7.24-6.98 (m, 2H), 6.07-5.98 (m, 1H), 5.82-5.60(min, 2H), 5.³¹ (m, 1H), 5.19-5.09 (m, 2H), 5.03-5.00 (m, 1H), 4.96-4.91(m, 2H), 4.83 (m, 1H), 4.69 (m, 1H), 4.51-4.37 (m, 2H), 4.34-4.29 (m,2H), 4.09-4.02 (m, 3H), 3.89-3.79 (m, 3H), 3.69 (m, 2H), 3.43-3.09 (m,2H), 2.39-2.07 (m, 2H), 2.02-2.00 (m, 1H), 1.83-1.52 (m, 2H), 1.43 (s,9H), 1.38-1.21 (m, 2H), 1.16 (m, 3H

³¹P (121.4 MHz, CDCl₃):

45.47 (s, ³¹P), 42.84 (s, ³¹P). LC/MS: M+1=650.

Alcohol (992 mg, 1.53 mmol) and DABCO (550 mg, 4.89 mmol) were dissolvedin toluene (8 mL). A toluene (8 mL) solution of brosylchloride (1.25 g,4.89 mmol) was added dropwise. The reaction mixture was stirred at r. t.for 3 h. The reaction was diluted with EtOAc and quenched with saturatedNaHCO₃. The two layers were separated and the organic layer was washedwith 0.5 N HCl, brine, dried with Na₂SO₄, filtered, and concentrated.The crude product was purified by combi-flash to give 764 mg ofbrosylate in 58% yield. ¹H NMR (300 MHz, CDCl₃):

7.81-7.

(m, 4H), 7.63 (m, 1H), 7.38 (m, 1H), 7.26-6.96 (m, 5H), 6.10-6.04 (m,1H), 5.80-5.70 (m, 1H), 5.28 (m, 1H), 5.18-4.87 (m, 6H), 4.64 (m, 1H),4.30-3.78 (m, 7H), 3.29-3.06 (m, 4H), 2.76-2.60 (m, 2H), 2.36-2.28 (m,2H), 2.22-2.05 (m, 2H), 1.74-1.56 (m, 3H), 1.43 (s, 9H), 1.39-1.10 (m,4H

³¹P (121.4 MHz, CDCl₃): d 46.3

(s, ³¹P), 43.32 (s, ³¹P). LC/MS: M+1=870.

Brosylate (760 mg, 0.87 mmol) was dissolved in CH₂Cl₂ (56 mL) anddegassed with N₂ for 20 minutes. Grubb's G1 (180 mg, 0.22 mmol) wasadded and degassed for an additional 20 minutes. The reaction mixturewas heated to 50° C. overnight and cooled to rt.Tris(hydroxymethyl)phosphine (1.35 g, 11 mmol) was added followed byaddition of TEA (3.1 mL, 22 mmol) and H₂O (10 mL). The reaction mixturewas heated to 50° C. for 4 h and then r.t. overnight. The two layerswere separated. The organic layer was washed with 0.5 N HCl, brine,dried with Na₂SO₄, and concentrated. The crude product was purified bycombi-flash to give 487 mg of cyclized olefin in 66% yield. ¹H NMR (300MHz, CDCl₃):

7.82-7.68 (m, 4H), 7.47-7.40 (m, 1H), 7.32-6.96 (m, 3H), 6.60-6.54 (m,1H), 5.66-4.98 (m, 6H), 4.46-3.97 (m, 8H), 3.84-3.74 (m, 1H), 3.44-3.13(m, 3H), 2.53-2.32 (m, 4H), 2.09-1.07 (m, 18H

³¹P (121.4 MHz, CDCl₃):

45.34 (s, ³¹P), 43.41 (s, ³¹P). LC/MS: M+1=842. A solution of olefin(798 mg, 0.95 mmol) and2-(2-isopropylamino-thiazol-4-yl)-7-methoxy-quinolin-4-ol (300 mg, 0.95mmol) in NMP (10 mL) was treated with Cs₂CO₃ (³¹0 mg, 0.95 mmol). Thereaction mixture was heated to 65° C. overnight and then cooled to rt.The reaction was diluted with EtOAc and washed with H₂O. The aqueouslayer was brought to pH=4 with 1 N HCl and extracted with EtOAc (2×).The combined organic layers were dried with Na₂SO₄ and concentrated. Thecrude product was purified by combi-flash to give 62.5 mg of desiredproduct in 72% yield. ¹H NMR (300 MHz, CDCl₃):

8.02-7.9

m, 1H), 7.47 (s, 1H), 7.34-6.96 (m, 7H), 5.69-5.60 (m, 1H), 5.40-5.25(m, 4H), 4.51-4.07 (m, 3H), 4.03-3.71 (m, 8H), 3.54-3.36 (m, 4H),3.16-3.05 (m, 2H), 2.85-2.68 (m, 2H), 2.40-1.07 (m, 28H); ³¹P (121.4MHz, CDCl₃):

45.15 s, ³¹P), 44.29 (s, ³¹P). LC/MS: M+1=919.

To a solution of product obtained above (625 mg, 0.68 mmol) in CH₃CN (10mL) at 0° C. was added iodotrimethylsilane (0.5 mL, 3.4 mmol). Thereaction mixture was stirred at 0° C. for 5 minutes. 2,6-Lutidine (0.48mL) was added and stirred for 1 h. TEA (2 mL) and MeOH (3 mL) were addedand stirred for 30 minutes. The mixture was concentrated, co-evaporatedwith toluene and CH₃CN, and dried for 20 minutes to give crude acid. Thecrude acid was dissolved in CH₃CN (5 mL). Saturated Na₂CO₃ in H₂O (5 mL)was added and stirred for 5 minutes. The THF solution of the freshlyprepared cyclopentylchloroformate was added. The reaction was completedwithin 0.5 h and concentrated. The residue was dissolved in EtOAc and1.0 N HCl was added to adjust pH=2. The two layers were separated andthe organic layer was concentrated. The crude product was purified byHPLC to give 327 mg of product 145 in 53% yield. ¹H NMR (300 MHz,CDCl₃):

8.89

s, 1H), 7.34 (d, J=9.5 Hz, 1H), 8.18 (s, 1H), 7.76-7.73 (m, 2H),7.39-7.02 (m, 5H), 5.84 (br s, 1H), 5.77 (dt, J=8.9 Hz, 9.1 Hz, 1H),5.29 (t, J=9.5 Hz, 1H), 4.86 (s, 1H), 4.72 (t, J=9.2 Hz, 1H), 4.38 (s,1H), 4.22-4.03 (m, 6H), 3.57-3.43 (m, 1H), 3.32-3.17 (m, 1H), 2.85-2.78(m, 2H), 2.66-2.58 (m, 1H), 2.³¹-2.23 (m, 1H), 1.94-1.82 (m, 3H),1.58-1.33 (m, 24H

³¹P (121.4 MHz, CD₃OD):

41.98 (s, ³¹P). LC/MS: M+1=903.

Example 146 Preparation of Compound 146

To a solution of 145 (30 mg, 0.033 mmol) in DME (1 mL)/H₂O (0.1 mL) wasadded p-tosylhydrazide (³¹ mg, 0.17 mmol) and NaOAc (19 mg, 0.23 mmol).The reaction mixture was heated to 95° C. for 2 h and cooled to rt. Afew drops of 3 N HCl was added to adjust pH=2. The crude product waspurified by HPLC to give 4 mg of acid 146. ¹H NMR (300 MHz, CD₃OD):

8.³¹ (d, J=9.4 Hz, 1H), 8.18 (s, 1H), 7.75 (s, 2H), 7.39-7.04 (m, 5H),5.84 (br s, 1H), 4.81-4.68 (m, 3H), 4.44 (s, 1H), 4.25-4.04 (m, 7H),3.47-3.23 (m, 2H), 2.84-2.77 (m, 2H), 2.55-2.52 (m, 2H), 1.95-1.33 (m,30H); ³¹P (121.4 MHz, CD₃OD):

3.74 s), ³¹P). LCMS: M+1=905.

Example 147 Preparation of Compound 147

Compound 147 was prepared as described for example 145 with thephosphinate in example 58. ¹H NMR (300 MHz, CD₃OD) d 8.32 (d, 1H, J=10Hz), 8.19 (s, 1H), 7.81-7.72 (m, 2H), 7.35-7.20 (m, 2H), 6.94 (t, 2H,J=8 Hz), 5.85 (br s, 1H), 5.74 (app q, 1H, J=8 Hz), 5.32 (t, 1H, J=10Hz), 4.74 (t, 1H, J=8 Hz), 4.40 (br s, 1H), 4.23-4.05 (m, 3H), 4.04 (s,3H), 3.58 (t, 1H, J=14 Hz), 3.36-3.22 (m, 1H), 2.90-2.77 (m, 1H),2.70-2.58 (m, 1H), 2.37-2.24 (m, 1H), 1.93-1.76 (m, 2H), 1.69-1.37 (m,17H), 1.34 (d, 6H, J=7 Hz); ¹⁹F NMR (282.6 MHz, CD₃OD) d −114.6; ³¹P(121.4 MHz, CD₃OD)

40.7; EI MS (m/z) 920.6 [MH⁺].

Example 148 Preparation of Compound 148

Compound was prepared as described for 146.

LC/MS: 923 (M⁺+1).

Example 149 Preparation of Compound 149

The brosylate from example 147 (1.49 g, 1.73 mmol) and thefluoro-quinoline (synthesis described below) (0.58 g, 1.73 mmol) weretaken up in NMP (18 mL) with cesium carbonate (0.57 g, 1.73 mmol). Thereaction was stirred at 60° C. for 15 h, cooled, and taken up in ethylacetate. The mixture was washed with bicarbonate solution and water,dried, concentrated and purified by flash chromatography to provide thedesired Boc-amine (0.942 g, 57%).

This Boc-amine (0.942 g, 0.098 mmol) was taken up in DCM (10 mL) and 4NHCl in dioxane (2.5 mL) was added. The mixture was stirred at room tempfor 1 h, then concentrated. The residue was taken up in acetonitrile (10mL) and water (10 mL). Cyclopentylchloroformate (5 equivalents) andsodium carbonate (0.125 g, 1.17 mmol) were added and the reactionstirred at room temp for 1.5 h. The reaction mixture was partitionedwith H₂O and ethyl acetate, washed with brine, dried (MgSO₄) andconcentrated. This residue was taken up in acetonitrile (10 mL) andsubjected to TMSI (0.70 mL, 4.93 mmol) for 15 minutes at which time2,6-lutidine (10 eq.) was added. The reaction was quenched withmethanol, concentrated and purified by HPLC to provide the desiredphosphinate compound 149 (533 mg, 58% 3 steps) ^(1H) NMR (300 MHz,CD₃OD): d 8.62 (s, 1H), 8.18 (m, 2H), 7.74 (s, 1H), 7.61 (m, 1H), 7.25(m, 1H), 6.93 (m, 2H), 5.79 (m, 2H), 5.36 (m, 1H), 4.76 (m, 2H), 4.38(m, 1H), 4.16 (m, 1H), 4.12 (s, 3H), 4.05 (m, 1H), 2.81 (m, 2H), 2.65(m, 1H), 2.32 (m, 1H), 1.86 (m, 1H), 1.60 (m, 22H), 1.37 (d, J=6.4 Hz,6H). ³¹P NMR (75 MHz, CD₃OD) d 40.807. LCMS: 940 (M−1).

The quinoline was prepared in the following manner:2-fluoro-3-methoxybenzoic acid (10 g, 58.8 mmol) and Hunig's base (12.3mL, 70.5 mmol) were taken up in toluene (50 mL) and tert-butanol (50 mL)and stirred over activated 4 angstrom molecular sieves for 1 h.Diphenylphosphorylazide (15.2 mL, 70.5 mmol) was added and the reactionwarmed to reflux overnight. The mixture was cooled, filtered andconcentrated. The residue was then taken up in ethyl acetate, washedwith water and brine, dried and concentrated to provide 15.6 g of crudematerial. This Boc-aniline was then subjected to 4N HCl in dioxane (260mL) for 1 h at room temp. The reaction was concentrated, then taken upin ethyl acetate, washed with sodium bicarbonate solution followed bybrine, dried, and concentrated to provide the aniline (1 g). This crudeaniline (10 g, 71 mmol)) was taken up in methanol (200 mL) anddimethylacetylene dicarboxylate (10.4 mL, 85 mmol) was added. Themixture was refluxed for 2 h, then concentrated and purified by columnchromatography (ethyl acateta/hexanes) to provide the desired product(11.64 g, 58%). This olefin (11.6 g) was taken up in diphenyl ether (80mL). A sand bath was prepared and warmed to 350° C. The reaction wasplaced in this warm sand bath and the internal temperature wasmonitored. When the internal temperature reached 240° C., a 5 minutetimer was started. After this time, the reaction was removed from thesand bath and allowed to cool. A brown solid precipitated out and thiswas filtered and washed extensively with diethyl ether to provide8-fluoro-4-hydroxy-7-methoxy-quinoline-2-carboxylic acid (5.5 g).

This methyl ester (3.95 g, 15.7 mmol) was taken up in THF (50 mL), water(50 mL), and methanol (50 mL) and LiOH (3.3 g, 79 mmol) was added. Thereaction was stirred at room temp for 1 h then acidified using HCl. Theproduct precipitated from solution and was then filtered and dried vialyophilization to provide the acid (3.57 g, 96%).

This acid (3.57 g, 15 mmol) was taken up in THF (150 mL) at zero 0° C.Triethylamine (4.6 mL, 33.1 mmol) and isobutylchloroformate (4.3 mL,33.1 mmol) was added and the mixture stirred for 1 h. At this time,diazomethane (2.2 equivalents) in ether solution was added (preparedfrom MNNG). The reaction was stirred at 0° C. for 30 minutes and warmedto room temp for 2 h. The mixture was then concentrated to provide thediazoketone quinoline with isobutyl carbonate protecting the hydroxyl.This diazoketone (15 mmol) was taken up in THF (100 mL) and cone HBr(8.5 mL, 75.3 mmol) was added at 0° C. The reaction was stirred for 1 h,then taken up in ethyl acetate, washed with bicarbonate solution, dried,and concentrated. The residue was taken up in isopropanol andisopropylthiourea (3.5 g, 30 mmol) was added. The reaction was heated to75° C. for 1 h, then cooled overnight. The orange solid was filtered anddried to provide the aminothiazolequinoline (1.7 g, 33%). ^(1H) NMR (300MHz, CD₃OD): d 8.62 (s, 1H), 8.18 (m, 2H), 7.74 (s, 1H), 7.61 (m, 1H),7.25 (m, 1H), 6.93 (m, 2H), 5.79 (m, 2H), 5.36 (m, 1H), 4.76 (m, 2H),4.38 (m, 1H), 4.16 (m, 1H), 4.12 (s, 3H), 4.05 (m, 1H), 2.81 (m, 2H),2.65 (m, 1H), 2.32 (m, 1H), 1.86 (m, 1H), 1.60 (m, 22H), 1.37 (d, J=6.4Hz, 6H). ³¹P NMR (75 MHz, CD₃OD) d 40.807. LC/MS: 940 (M−1).

Example 150 Preparation of Compound 150

Compound 149 (528 mg, 0.56 mmol) was taken up in DME (5 mL) and water(0.5 mL). Sodium acetate (0.69 g, 8.43 mmol) and tosyl hydrazide (0.785g, 4.21 mmol) were added and the reaction heated at 95° C. The reactionwas monitored by LCMS and determined to be complete after 5 h at whichtime it was cooled to 0° C. and HCl (1.4 mL of 6N solution) was addedand the reaction was concentrated. The residue purified by HPLC toprovide the desired saturated product compound 150 (390 mg, 74%). ¹H NMR(300 MHz, CD₃OD) d 8.16 (m, 2H), 7.72 (s, 1H), 7.64 (m, 1H), 7.27 (m,1H), 6.95 (m, 2H), 5.81 (br s, 1H), 4.75 (m, 2H), 4.47 (m, 1H), 4.28 (m,1H), 4.12 (s, m, 4H), 4.04 (m, 1H), 2.80 (m, 1H), 2.59 (m, 1H), 1.98 (m,1H), 1.82 (m, 2H), 1.37 (m, ³¹H). ³¹P NMR (121.4 MHz, CD₃OD) d 42.381.LCMS: 942 (M+1).

Example 151 Preparation of Compound 151

Step 1. To a mixture of intermediate VIII (22.03 g, 60.12 mmol) and(1-amino-2-vinyl-cyclopropyl)-(2,6-difluoro-benzyl)-phosphinic acidethyl ester (described in Example 58, 12.97 g, 40.48 mmol) in toluene(160 mL) was added sodium 2-ethylhexanoate (10.01 g, 60.26 mmol) and H₂O(240 mL). The reaction mixture was heated to 80° C. and stirred for 48h. The reaction mixture was cooled to room temperature, diluted withEtOAc (400 mL), and separated. The aqueous layer was extracted withEtOAc (200 mL). The combined organic layers were washed with 1 N HCl(400 mL), NaHCO₃ (300 mL), brine (400 mL), dried with Na₂SO₄, andconcentrated in vacuo. The coupling product (23.25 g) was obtained ascrude product. LCMS (M+1): 667.97

Step 2. The product obtained above (23.25 g, 34.82 mmol) and DABCO (6.26g, 55.79 mmol) were dissolved in toluene (55 mL). A toluene (15 mL)solution of 4-bromobenzenesulfonyl chloride (12.48 g, 48.9 mmol) wasadded. The reaction mixture was stirred at r. t. overnight whereuponadditional 4-bromo-benzenesulfonyl chloride (7.14 g, 39.12 mmol) andDABCO (3.13 g, 27.9 mmol) were added. The reaction was stirred for 3 h.The reaction mixture was diluted with EtOAc (350 mL) and 0.5 N HCl (400mL) was added. The two layers were separated. The organic layers werewashed with brine (400 mL), dried with Na₂SO₄, filtered, andconcentrated in vacuo. The crude product was purified by silica gelchromatography (EtOAc-hexanes) to give 20.38 g of brosylate. LCMS (M+1):887.75

Step 3. Brosylate (11.96 g, 13.49 mmol) was dissolved in CH₂Cl₂ (1.33 L)and the solution was degassed for 30 min. The solution was heated to 40°C. and Grubb's G1 catalyst (2.78 g, 3.38 mmol) was added. The reactionwas heated to 45° C. and stirred overnight. Additional Grubb's G1catalyst (567 mg) was added and stirred for 7 h at 45° C.Trishydroxymethylphosphine (25.24 g, 0.17 mol), TEA (57 mL, 0.34 mol),and H₂O (200 mL) were added and the reaction mixture was refluxedovernight whereupon it was cooled to rt and the two layers wereseparated. The organic layer was washed with H₂O (2×200 mL) and brine(400 mL), dried with Na₂SO₄, and concentrated in vacuo. The crudeproduct was purified by silica gel chromatography (EtOAc-hexanes) togive 5.79 g of macrocyclic olefin in 50% yield. LCMS (M+1): 857.88

Step 4. Macrocyclic olefin (5.78 g, 6.74 mmol) and8-chloro-4-hydroxy-7-methoxy-quinoline-2-carboxylic acid methyl ester(2.17 g, 8.10 mmol) were dissolved in NMP (68 mL) and Cs₂CO₃ (2.21 g,6.77 mmol) was added. The reaction mixture was heated to 70° C. andstirred for 6 h. Additional Cs₂CO₃ (219 mg, 0.68 mmol) was added andstirred at 70° C. for 5 h. The reaction mixture was cooled to r.t. andstirred overnight. The mixture was poured into H₂O/brine (900 mL/100 mL)and extracted with EtOAc (3×250 mL). The organic layer was washed with2% LiCl (300 mL), NaHCO₃ (300 mL), brine (300 mL), dried with Na₂SO₄,filtered, and concentrated. The crude product was purified by silica gelchromatography (EtOAc-hexanes) to give 4.11 g of ester in 69% yield.LCMS (M+1): 889.14

Step 5. To a solution of ester obtained above (4.11 g, 4.62 mmol) inCH₂Cl₂ (23 mL) was added 4 N HCl in 1,4-dioxane (23 mL). The reactionmixture was stirred at r.t for 1.5 h and concentrated in vacuo. Thecrude HCl salt was dissolved in CH₃CN (46 mL)/saturated Na₂CO₃ (46 mL)and a solution of cyclopentylchloroformate in THF (46 mL) was added. Thereaction was completed within 20 min. The organic was decanted from thesolid that precipitated and concentrated in vacuo after washing thesolid with CH₃CN and CH₂Cl₂. The solid that precipitated was dissolvedin H₂O (250 mL) and extracted with CH₂Cl₂ (2×125 mL). The combinedorganic layers were washed with brine, dried with Na₂SO₄, andconcentrated in vacuo. The crude product was purified by silica gelchromatography (EtOAc-hexanes) to give 4.00 g of cyclopentyl carbamate.LCMS (M+1): 901.13

Step 6. Cyclopentyl carbamate (4.39 g, 4.87 mmol) was dissolved in THF(48 mL)/H₂O (48 mL) and cooled to 0° C. NaOH (200 mg, 5.0 mmol) wasadded. The reaction mixture was stirred at 0° C. for 40 min andadditional NaOH (200 mg, 5.0 mmol) was added. The reaction was stirredfor 15 min. The reaction mixture was diluted with H₂O (200 mL),acidified to pH=2 with 1 N HCl, and extracted with EtOAc (3×300 mL). Thecombined organic layers were dried with Na₂SO₄, filtered, concentrated,and dried under vacuum to give 4.26 g of acid in 98% yield.

Step 7. To a solution of acid (2.20 g, 2.48 mmol) in THF (25 mL) at 0°C. was added TEA (0.38 mL, 2.73 mmol) and stirred for 5 min.Isobutylchloroformate (0.36 mL, 2.75 mmol) was added dropwise and thereaction mixture was stirred for 40 min at 0° C. An ether solution ofdiazomethane (5 mL, 5 mmol) was added and the reaction mixture waswarmed to r.t. and stirred for 2 h. The mixture was concentrated and theresidue was dissolved in EtOAc (400 mL). The EtOAc layer was washed withsaturated NaHCO₃ (150 mL), H₂O (150 mL), brine (150 mL), dried withNa₂SO₄, and concentrated to give 2.22 g of crude diazoketone product.LCMS (M+1): 911.33

Step 8. Diazoketone (2.22 g, 2.44 mmol) was dissolved in THF (25 mL) andcooled to 0° C., whereupon aqueous HBr (1.38 mL, 48%, 12.2 mmol) wasadded dropwise and the reaction was stirred for 1.5 h. The reactionmixture was diluted with EtOAc (400 mL) and washed with NaHCO₃. Theaqueous layer was extracted with EtOAc (150 mL). The combined organiclayers were dried with Na₂SO₄, filtered, concentrated, and dried undervacuum to give 2.36 g of α-bromoketone. LCMS (M+1): 965.01

Step 9. A mixture of α-bromoketone (2.36 g, 2.44 mmol) andisopropyl-thiourea (580 mg, 4.91 mol) in 2-propanol (25 mL) was heatedto 75° C. and stirred for 1.5 h. The reaction mixture was cooled to r.t. and concentrated. The residue was dissolved in EtOAc (300 mL) andwashed with NaHCO₃ (350 mL). The aqueous layer was extracted with EtOAc(200 mL). The combined organic layers were washed with brine (200 mL),dried with Na₂SO₄, and concentrated to give 2.64 g of ester product.LCMS (M+1): 983.31

Step 10. Ester (2.64 g, 2.68 mmol) was dissolved in CH₃CN (18 mL) andcooled to 0° C. Iodotrimethylsilane (1.95 mL, 13.7 mmol) was addeddropwise. The reaction mixture was warmed to r.t. and stirred for 35min. 2,6-lutidine (4 mL) and MeOH (4 mL) were added and the reactionmixture was concentrated. The crude product was purified by HPLC to give1.78 g of Compound 151. ¹H NMR (300 MHz, CDCl₃) d 8.38 (d, J=9.3 Hz,1H), 8.30 (s, 1H), 7.60 (d, J=9.3 Hz, 1H), 7.32-7.19 (m, 1H), 6.99-6.87(m, 2H), 5.84 (bs, 1H), 5.73 (bq, J=8.9 Hz, 1H), 5.32 (dd, J=9.9, 9.9Hz, 1H), 4.96-4.82 (m, 1H), 4.80-4.71 (m, 1H), 4.27 (bs, 1H), 4.20-3.78(m, 3H), 4.16 (s, 3H), 3.66-3.52 (m, 1H), 3.36-3.22 (m, 1H), 2.92-2.73(m, 2H), 2.73-2.59 (m, 1H), 2.35-2.22 (m, 1H), 1.93-1.71 (m, 2H)1.71-1.20 (m, 17H) 1.38 (d, J=6.6 Hz, 6H). ³¹P NMR (121.4 MHz, CD₃OD)

40.5; LCMS (M+1): 955.43.

Example 152 Preparation of Compound 152

Step 1. Intermediate XI (17.42 g, 28.30 mmol) was dissolved in THF (136mL) and cooled to 0° C. To the solution was added N-methylmorpholine(4.7 mL, 42.7 mmol). After 10 min at 0° C., i-butylchloroformate (4.05mL, 30.96 mmol) was added dropwise. After an additional 1 h,(1-amino-2-vinyl-cyclopropyl)-(2,6-difluoro-benzyl)-phosphinic acidethyl ester (described in Example 58, 8.94 g, 29.70 mmol) was slowlyadded as a soln in THF (20 mL). The suspension was warmed to rt andafter 2 h it was partitioned between H₂O (400 mL) and ethylacetate (200mL). The aqueous layer was extracted with ethylacetate (200 mL×2) andthe combined organic layers were washed with HCl (1N, 225 mL) and H₂O(200 mL). The acid wash and aqueous wash were combined andback-extracted with ethylacetate (175 mL×2, 100 mL×2). The combinedorganic layers were washed with brine (400 mL), dried over Na₂SO₄, andconcentrated in vacuo providing 25.06 g of diene product in 98.5% crudeyield. LCMS (M+1): 898.06

Step 2. The crude diene product (12.91 g, 14.36 mmol) was dissolved inCH₂Cl₂ (1440 mL) and the solution was degassed for 30 min. The solutionwas heated to 40° C. and Grubb's G1 catalyst (2.95 g, 3.59 mmol) wasadded. The reaction was refluxed for 17 h whereupontris-hydroxymethylphosphine (22.3 g, 18.0 mmol), TEA (50 mL, 35.9 mmol),and H₂O (400 mL) were added and the reaction mixture was heated toreflux for an additional 16 h. The reaction mixture was cooled to r.t.and the two layers were separated. The organic layer was washed with H₂O(400 mL) and brine (300 mL), dried over MgSO₄, and concentrated. Thecrude residue was purified by silica-gel chromatography to afford 8.30 gof macrocyclic olefin product in 66% yield. LCMS (M+1): 870.09.

Step 3. The macrocyclic olefin (7.34 g, 8.42 mmol) was dissolved inethylacetate (105 mL) and rhodium on alumina (5% wt, 2.945 g, 0.40 wt %)was added. The system was evacuated and flushed with H₂ (1 atm, 3×). Tothe system, after 3 h, was added more rhodium on alumina (5% wt, 842 mg,0.10 wt %) and it evacuated and flushed with H₂ (1 atm, 3×). After anadditional 1 h the suspension was filtered and coned in vacuo providing6.49 g of reduced macrocycle in 88% crude yield. LCMS (M+1): 872.04.

Step 4. The brosylate macrocycle (6.49 g, 7.67 mmol) was dissolved inN-methylpyrrolidinone (25.0 mL) and8-chloro-2-(2-isopropylamino-thiazol-4-yl)-7-methoxy-quinolin-4-ol(2.564 g, 7.33 mmol) followed by Cs₂CO₃ (4.40 g, 13.50 mmol) were added.The mixture was heated to 65° C. for 6 h then diluted with ethylacetate(200 mL) and washed with LiCl (5%, 250 mL). The aqueous layer wasextracted with ethylacetate (100 mL×2) and the combined organic layerswere washed with brine (150 mL), dried over Na₂SO₄/MgSO₄, andconcentrated in vacuo. The crude residue was purified via silica-gelchromatography (ethylacetate-methanol) affording 4.39 g of aminothiazoleproduct in 58% yield. LCMS (M+1): 985.28.

Step 5. Phosphinate ester (23.7 g, 24.05 mmol) was dissolved in CH₃CN(240 mL) and cooled to 0° C. Iodotrimethylsilane (17.4 mL, 122.3 mmol)was added at a fast drop-wise pace followed by, after 10 min,2,6-lutidine (17.0 mL, 146.4 mmol). The reaction mixture was slowlywarmed to r.t. and stirred for 1 h then cooled back down to 0° C. and2,6-Lutidine (11.1 mL, 95.6 mmol) followed by MeOH (24 mL) were added.The solution was concentrated in vacuo and the crude residue waspurified by HPLC to afford 12.68 g of Compound 152 in 55% yield. ¹H NMR(300 MHz, CDCl₃) d 8.35 (d, J=9.3 Hz, 1H), 8.28 (s, 1H), 7.85 (s, 1H

7.64 (d, J=9.6 Hz, 1H), 7.35-7.22 (m, 1H), 7.02-6.89 (m, 2H), 5.85 (bs,1H), 4.82-4.71 (m, 2H), 4.33 (bs, 1H), 4.28-3.99 (m, 3H), 4.16 (s, 3H),3.57-3.28 (m, 2H), 2.90-2.7

m, 1H), 2.63-2.50 (m, 1H), 2.08-1.91 (m, 1H), 1.91-170 (m, 2H),1.70-1.13 (m, 22H), 1.37 (d, J=6.9 Hz, 6H); ³¹P NMR (121.4 MHz, CD₃OD)

42.4; LCMS (M+1): 957.35.

Example 153 Preparation of Compound 153

The brosylate from example 147 (603.2 mg, 0.70 mmol) and8-bromo-4-hydroxy-7-methoxy-quinoline-2-carboxylic acid methyl ester(263.4 mg, 0.84 mmol) were dissolved in NMP (7.0 mL) and Cs₂CO₃ (251.5mg, 0.77 mmol) was added. The reaction mixture was heated to 70° C. andstirred overnight. The reaction mixture was cooled to r.t. and pouredinto H₂O (175 mL)/brine (25 mL) and extracted with EtOAc (3×100 mL). Theorganic layer was washed with 2% LiCl (125 mL), NaHCO₃ (150 mL), brine(100 mL), dried with Na₂SO₄, filtered, and concentrated to give 655 mgof crude product. LCMS (M+1): 934.94.

To a solution of product obtained above (655 mg, 0.70 mmol) in CH₂Cl₂(3.6 mL) was added 4 N HCl in 1,4-dioxane (3.4 mL). The reaction mixturewas stirred at r.t for 1 h and concentrated in vacuo. The crude HCl saltwas dissolved in CH₃CN (7 mL)/saturated Na₂CO₃ (7 mL) and a solution ofcyclopentylchloroformate in THF was added. The reaction was completedwithin 20 min. The organic was decanted from the Na₂CO₃ thatprecipitated and concentrated in vacuo after washing the solid withCH₃CN and CH₂Cl₂. The solid Na₂CO₃ was dissolved in H₂O (75 mL) andextracted with CH₂Cl₂ (2×75 mL). The combined organic layers were washedwith brine (75 mL), dried with Na₂SO₄, and concentrated. The crudeproduct was purified by combi-flash to give 473 mg of cyclopentylcarbamate in 71% yield. LCMS (M+1): 945.10.

Methyl ester obtained above (473 mg, 0.5 mmol) was dissolved in THF (1.6mL)/H₂O (1.7 mL) and cooled to 0° C. LiOH (60.4 mg, 2.52 mmol) in H₂O(1.7 mL) was added. The reaction mixture was stirred at 0° C. for 40 minand additional NaOH (200 mg, 5.0 mmol) was added. The reaction wasstirred for 15 min. The reaction mixture was diluted with H₂O (20 mL),acidified to pH=1 with 1 N HCl, and extracted with EtOAc (3×25 mL). Thecombined organic layers were dried with Na₂SO₄, filtered, concentrated,and dried under vacuum to give acid.

To a solution of acid (466 mg, 0.50 mmol) in THF (5 mL) at 0° C. wasadded TEA (77 μL, 0.55 mmol) and stirred for 5 min.Isobutylchloroformate (72 μL, 0.55 mmol) was added dropwise and thereaction mixture was stirred for 1 h at 0° C. An ether solution ofdiazomethane (2.70 mL, 1.08 mmol) was added and the reaction mixture waswarmed to r.t. and stirred for 2 h. The mixture was concentrated and theresidue was dissolved in EtOAc (75 mL). The EtOAc layer was washed withsaturated NaHCO₃ (60 mL), H₂O (50 mL), brine (50 mL), dried with Na₂SO₄,and concentrated to give crude product.

Diazoketone crude (478 mg, 0.5 mmol) was dissolved in THF (5 mL) andcooled to 0° C., HBr (0.29 mL, 2.52 mmol) was added dropwise and thereaction was stirred for 15 min. The reaction mixture was diluted withEtOAc (200 mL) and washed with NaHCO₃ (75 mL). The aqueous layer wasextracted with EtOAc (20 mL). The combined organic layers were driedwith Na₂SO₄, filtered, concentrated, and dried under vacuum to give403.4 mg of bromoketone. LCMS (M+1): 1008.91.

A mixture of bromoketone (403.4 mg, 0.4 mmol) and isopropyl-thiourea(94.4 mg, 0.8 mol) in 2-propanol (25 mL) was heated to 75° C. andstirred for 0.5 h. The reaction mixture was cooled to r. t. andconcentrated. The residue was dissolved in CH₂Cl₂ (150 mL) and washedwith NaHCO₃ (30 mL). The aqueous layer was extracted with CH₂Cl₂ (150mL). The combined organic layers were washed with brine (50 mL), driedwith Na₂SO₄, and concentrated to give 424.1 mg of desired product. LCMS(M+1): 1029.17.

Ester obtained above (424.1 mg, 0.41 mmol) was dissolved in CH₃CN (4.1mL) and cooled to 0° C. Iodotrimethylsilane (0.3 mL, 2.07 mmol) wasadded dropwise. The reaction mixture was warmed to r.t. and stirred for20 min and cooled to 0° C. 2,6-lutidine (0.5 mL) and MeOH (0.5 mL) wereadded and the reaction mixture was concentrated. The crude product waspurified by HPLC to give 232.1 mg of compound 153 in 46% yield. ¹H NMR(300 MHz, CDCl₃) d: 8.43 (d, J=9.0 Hz, 1H), 8.33 (s, 1H), 7.90 (s, 1H),7.60 (d, J=9.3 Hz, 1H), 7.35-7.21 (m, 1H), 6.95 (dd, J=7.8, 7.5 Hz, 2H)5.88 (s, 1H), 5.84-5.71 (m, 1H), 5.34 (dd, J=9.3, 9.0 Hz, 1H), 4.77 (dd,J=8.4, 7.5 Hz, 1H), 4.24-4.01 (m, 7H), 3.60 (t, J=15.3 Hz, 1H), 3.32 (t,J=15.3 Hz, 1H), 2.95-2.75 (m, 2H), 2.75-2.58 (m, 1H), 2.37-2.23 (m, 1H),1.96-1.61 (m, 2H), 1.61-1.15 (m, 17H) 1.38 (d, J=6.6 Hz, 6H); ³¹P (121.4MHz, CD₃OD)

40.9; LCMS (M+): 1001.20.

Example 154 Preparation of Compound 154

A mixture of 153 (221.3 mg, 0.22 mmol), sodium acetate (274.1 mg, 3.34mmol), and p-tosylhydrazine (³¹0.7 mg, 1.67 mmol) in DME (2 mL) and H₂O(0.2 mL) was heated to 95° C. for 1.5 h. The reaction mixture was cooledto r.t. and treated with 4 N HCl (0.8 mL). The crude product waspurified by HPLC to give 101 mg of 154 in 47% yield. ¹H NMR (300 MHz,CDCl₃) d 8.40 (d, J=9.3 Hz, 1H), 8.29 (s, 1H), 7.88 (s, 1H), 7.63 (d,J=9.6 Hz, 1H), 7.33-7.21 (m, 1H), 6.96 (t, J=7.8 Hz, 2H) 5.86 (bs, 1H),4.83-4.70 (m, 1H), 4.³¹-4.00 (m, 5H), 4.17 (s, 3H), 3.57-3.47 (m, 1H),3.47-3.33 (m, 1H), 2.90-2.78 (m, 1H), 2.64-2.52 (m, 1H), 2.06-1.92 (m,1H), 1.90-1.69 (m, 2H), 1.69-1.12 (m, 22H), 1.37 (d, J=6.6 Hz, 6H); ³¹P(121.4 MHz, CD₃OD)

42.2.

Example 155 Preparation of Compound 155

A solution of macrocyclic phosphinate (example 147) (212.6 mg, 0.25mmol) and2-(2-isopropylamino-thiazol-4-yl)-7-methoxy-8-methyl-quinolin-4-ol (82.0mg, 0.25 mmol) in NMP (3 mL) was treated with Cs₂CO₃ (81.3 mg, 0.25mmol). The reaction mixture was heated to 70° C. overnight and thencooled to rt. The reaction was diluted with 5% LiCl (10 mL) andextracted with EtOAc (3×10 mL). The combined organic layers were washedwith brine, dried with Na₂SO₄, and concentrated. The crude product waspurified by combi-flash to give 166.1 mg of desired product in 70%yield.

To a solution of compound obtained above (1.383 g, 1.45 mmol) in CH₂Cl₂(7.5 mL) was added 4 N HCl in 1,4-dioxane (7.5 mL, 30 mmol). Thereaction mixture was stirred at r.t. for 2 h, concentrated, dried undervacuum for 20 minutes, and then dissolved in CH₃CN (15 mL). SaturatedNaHCO₃ in H₂O (15 mL) was added and stirred for 5 minutes. The freshlyprepared cyclopentylchloroformate (7.7 mmol) in THF (15 mL) was added.The reaction was completed within 1 h. The solvent was removed onrotavap and the residue was diluted with EtOAc. The mixture was broughtto pH=2 with 1 N HCl and the two layers were separated. The organiclayers were washed with brine, dried with Na₂SO₄, filtered, andconcentrated. Cyclopentanyl carbamate (1.21 g) was obtained as crudeproduct.

To a solution of cyclopentanyl carbamate (194.9 mg, 0.20 mmol) in CH₃CN(2 mL) at 0° C. was added 5 equivalents of iodotrimethylsilane. Thereaction mixture was stirred at 0° C. for 35 minutes. 2,6-lutidine (0.4mL) was added and stirred for 5 minutes. MeOH (0.4 mL) was added andstirred for 20 minutes. The mixture was concentrated and the crude acidwas purified by HPLC to give 97.4 mg of acid 155 in 51% yield.LC/MS=935.40 (M⁺+1)

Example 156 Preparation of Compound 156

To a solution of 155 (46.8 mg, 0.05 mmol) in DME (0.5 mL)/H₂O (0.05 mL)was added p-tosylhydrazide (69.7 mg, 0.80 mmol) and NaOAc (61.8 mg, 0.75mmol). The reaction mixture was heated to 95° C. for 2.5 h and cooled tort. A few drops of 3 N HCl was added to adjust pH=2. The crude productwas purified by HPLC to give 20.6 mg of acid 156 in 44% yield.LC/MS=937.33 (M⁺+1)

Example 157 Preparation of Compound 157

The acid XI (3.09 g, 5.03 mmol) and NMM (0.78 mL, 7.06 mmol) weredissolved in THF (40 mL) and cooled to 0° C. Isobutylchloroformate (0.69mL, 5.3 mmol) was added dropwise and stirred at 0° C. for 1 h.(1-Amino-2-vinyl-cyclopropyl)-(2,3,6-trifluoro-benzyl)-phosphinic acidethyl ester (prepared as described in example 68 with DMS) (1.75 g, 5.48mmol) was added slowly and the mixture was stirred at r.t. for 1.5 h.The reaction was diluted with EtOAc and washed with 1 N HCl, saturatedNaHCO₃/brine, dried with Na₂SO₄ and concentrated. The crude product waspurified by combi-flash to give 2.7 g of compound diene in 59% yield.

Diene compound (2.7 g, 2.94 mmol) was dissolved in CH₂Cl₂ (300 mL) anddegassed with N₂ for 20 minutes. Grubb's G1 (970 mg, 1.18 mmol) wasadded and degassed for an additional 20 minutes. The reaction mixturewas heated to 45° C. overnight and cooled to rt.Tris(hydroxymethyl)phosphine (9 g) was added followed by addition of TEA(20 mL) and H₂O (40 mL). The reaction mixture was heated to 50° C.overnight. The two layers were separated. The organic layer was washedwith 0.5 N HCl, brine, dried with Na₂SO₄, and concentrated. The crudeproduct was purified by combi-flash to give 2.01 g of macrocycliccompound in 77% yield.

A solution of macrocyclic compound (2.0 g, 2.25 mmol) and8-chloro-2-(2-isopropylamino-thiazol-4-yl)-7-methoxy-quinolin-4-ol (786mg, 2.25 mmol) in NMP (11 mL) was treated with Cs₂CO₃ (880 mg, 2.7mmol). The reaction mixture was heated to 60° C. for 1 h. AdditionalCs₂CO₃ (1.13 g, 3.47 mmol) was added and stirred at 60° C. for 1.5 h.The temperature was lowered to 40° C. overnight and then cooled to rt.The reaction was diluted with EtOAc and washed with 5% LiCl (3×) andbrine. The organic layer was dried with Na₂SO₄ and concentrated. Thecrude product was directly used for next step reaction.

The residue (2.07 g, 2.07 mmol) was dissolved in CH₃CN (20 mL) at 0° C.To this mixture was added iodotrimethylsilane (1.48 mL, 10.39 mmol). Thereaction mixture was stirred at 0° C. for 5 minutes. 2,6-Lutidine (1.44mL, 12.46 mmol) was added and stirred for 1.5 h. MeOH was added andstirred for 30 minutes. The mixture was concentrated and re-dissolved inminimal MeOH and divided into two portions. One portion was purified byHPLC to give 404 mg of 157. LC/MS=973 (M⁺+1)

Example 157X Preparation of Compound 157X

Compound 157 was reduced to give 157X as described for example 156.LC/MS=975 (M⁺+1)

Example 158 Preparation of Compound 158

To a solution of(1-benzyloxycarbonylamino-2-vinyl-cyclopropyl)-(3-chloro-2,6-difluoro-benzyl)-phosphinicacid ester (example 65) (8 g, 17.1 mmol) in TFA (46 mL, 614 mmol) at 0°C. was added DMS (12.1 mL, 164.2 mmol). The reaction mixture was stirredat r.t. overnight. The reaction mixture was poured into ice cold 4 N HCl(500 mL) and extracted with 1/1 iPrOH/heptane (500 mL). The organiclayers were washed with 4 N HCl (5×500 mL). The combined aqueous layerswere brought to pH=10 in a cold bath. The aqueous layer was extractedwith EtOAc (5×500 mL). The organic layers were washed with brine, driedwith Na₂SO₄, and concentrated to give 4.1 g of amine in 66% yield.

The acid from intermediate X (8.4 g, 13.86 mmol) and amine obtainedabove (3.9 g, 11.55 mmol) were dissolved in DMF (100 mL). HATU (10.97 g,28.88 mmol) and NMM (5.9 g, 57.75 mmol) were added and the mixture wasstirred at r.t. for 2 h. The reaction was diluted with EtOAc and washedwith 20% LiCl (2×500 mL). The organic layer was washed with aqueousNH₄Cl (500 mL), dried with Na₂SO₄, and concentrated. The crude productwas purified by combi-flash to give 8.2 g of tripeptide. ¹H NMR (300MHz, CDCl₃

d 7.74 (m, 4H), 7.24 (m, 1H), 6.83 (m, 1H), 6.07 (m, 1H), 5.78 (m, 2H),5.25 (m, 1H), 5.17 (m, 1H), 4.98 (m, 1H), 4.67 (d, 1H), 4.38 (m, 2H),4.18 (m, 2H), 4.12 (m, 2H), 3.83 (m, 2H), 3.65 (m, 2H), 3.32 (m, 3H),2.24 (m, 3H), 2.04 (m, 4H), 1.80 (m, 2H), 1.63 (m, 1H), 1.51 (m, 2H),1.42 (m, 9H), 1.25 (m, 3H). ³¹P (121.4 MHz, CDCl₃):

45.110, 44.254 diastereomers.

Tripeptide (5.0 g, 5.43 mmol) was dissolved in CH₂Cl₂ (600 mL) anddegassed with N₂ for 30 minutes. Grubb's G1 (1.34 g, 1.63 mmol) wasadded and degassed for an additional 30 minutes. The reaction mixturewas heated to 45° C. overnight and cooled to rt.Tris(hydroxymethyl)phosphine (9.9 g, 80.1 mmol) was added and stirredfor 20 minutes. TEA (16.5 g, 162.9 mmol) was added and stirred for 20minutes followed by addition of H₂O (60 mL). The reaction mixture washeated to 50° C. for 4 h and then r.t. overnight. The two layers wereseparated. The organic layer was washed with 0.5 N HCl, brine, driedwith Na₂SO₄, and concentrated. The crude product was purified bycombi-flash to give 2.1 g of cyclized compound. ¹H NMR (300 MHz, CDOD₃

d 7.87 (m, 4H), 7.40 (m, 1H), 7.03 (m, 1H), 6.75 (m, 1H), 5.78 (m, 1H),5.60 (m, 1H), 5.40 (m, 1H), 5.11 (m, 1H), 4.90 (m, 1H), 4.37 (m, 3H),3.83 (m, 2H), 3.75 (m, 2H), 3.51 (m, 1H) 3.22 (m, 2H), 2.24 (m, 3H),2.04 (m, 4H), 1.80 (m, 2H), 1.63 (m, 1H), 1.51 (m, 2H), 1.42 (m, 9H),1.25 (m, 3H). ³¹P (121.4 MHz, CDOD₃):

45.978, 45.613 diastereomers.

A solution of cyclic phosphinate (2.1 g, 2.35 mmol) and8-chloro-2-(2-isopropylamino-thiazol-4-yl)-7-methoxy-quinolin-4-ol (820mg, 2.35 mmol) in NMP (50 mL) was treated with Cs₂CO₃ (1.53 g, 4.7mmol). The reaction mixture was heated to 80° C. for 6 h and then cooledto rt. The reaction was diluted with EtOAc and washed with H₂O. Theaqueous layer was brought to pH=4 with 1 N HCl and extracted with 10%MeOH/EtOAc (2×200 mL). The combined organic layers were concentrated.The crude material was dissolved in CH₂Cl₂ (1 L), dried with Na₂SO₄ andconcentrated. The crude product was dried in vacuo and carried on fornext step reaction.

To a solution of crude product obtained above (1.5 g, 1.5 mmol) inCH₂Cl₂ (5 mL) was added 4 N HCl in 1,4-dioxane (5 mL, 20 mmol). Thereaction mixture was stirred at r.t. for 0.5 h, concentrated, driedunder vacuum for 20 minutes, and then dissolved in CH₃CN (20 mL). Na₂CO₃(1.5 g, 18 mmol) in H₂O (30 mL) was added and stirred for 5 minutes. ATHF solution of the freshly prepared cyclopentylchloroformate (1.11 g,7.5 mmol) was added. The reaction was completed within 1 h. The solventwas removed on rotavap and the residue was diluted with EtOAc. Themixture was brought to pH=2 with 1 N HCl and the two layers wereseparated. The organic layers were washed with brine, dried with Na₂SO₄,filtered, and concentrated. The crude product was dried under vacuoovernight and dissolved in CH₃CN (3 mL)/CH₂Cl₂ (1 mL). The reactionmixture was cooled to 0° C. and iodotrimethylsilane (1.1 mL, 7.5 mmol)was added. The reaction mixture was warmed to r.t and stirred for 30minutes and cooled to 0° C. 2,6-Lutidine (1 mL) and MeOH (1 mL) wereadded and stirred for 10 minutes. The mixture was concentrated and thecrude product was purified by HPLC to give 560 mg of product 158. ¹H NMR(300 MHz, CD₃OD): d 8.91 (s, 1H), 8.36 (d, J=9.0 Hz, 2H), 8.30 (s, 1H),7.85 (s, 1H), 7.65 (d, J=9.6 Hz, 2H), 7.39 (m, 1H), 7.00 (t, J=9.0 Hz,1H), 5.85 (s, 1H), 5.77 (m, 1H), 5.33 (m, 1H), 4.78 (m, 1H), 4.38 (m,1H), 4.22 (m, 2H), 4.07 (s, 3H), 3.75 (m, 1H), 3.38 (m, 2H), 2.85 (m,1H), 2.57 (m, 1H) 1.98-1.37 (m, 2H), 1.33 (m, 6H). ³¹P NMR (121.4 MHz,CD₃OD):

40.231; LC/MS=989 (M⁺+1)

Example 159 Preparation of Compound 159

To a solution of 158 (771 mg, 0.79 mmol) in DME (5 mL)/H₂O (0.4 mL) wasadded p-tosylhydrazide (737 mg, 3.96 mmol) and NaOAc (650 mg, 7.93mmol). The reaction mixture was heated to 95° C. for 1.5 h and cooled tort. A few drops of 3 N HCl was added to adjust pH=2. The crude productwas purified by HPLC to give 587 mg of acid 159 in 76% yield. ¹H NMR(300 MHz, CD₃OD): d 8.89 (s, 1H), 8.39 (d, J=9.5 Hz, 2H), 8.25 (s, 1H),7.82 (s, 1H), 7.60 (d, J=9.5 Hz, 2H), 7.40 (m, 1H), 7.01 (m, 1H), 5.84(s, 1H), 4.75 (m, 1H), 4.38 (m, 1H), 4.22 (m, 2H), 4.07 (s, 3H), 3.75(m, 1H), 3.38 (m, 2H), 2.83 (m, 1H), 2.58 (m, 1H), 2.31 (m, 1H) 1.82 (m,1H), 1.58 (m, 23H), 1.35 (m, 6H). ³¹P (121.4 MHz, CD₃OD):

41.136; LC/MS=991 (M⁺+1)

Example 160 Preparation of Compound 160

Step 1. A solution of N-Boc-L-serine (10.26 g, 50 mmol) in DMSO (200 mL)was treated with NaH (4 g, 100 mmol). The mixture was stirred at rt for1 h and 5-bromo 1-pentene (6 mL, 50 mmol) was added. An additional NaH(4 g, 100 mmol) was added and the reaction mixture was stirredovernight. The reaction was diluted with EtOAc (200 mL) and quenchedslowly with H₂O. EtOAc (200 mL) was added and 1 N HCl was added toadjust pH=2. The two layers were separated. The organic layer was washedwith H₂O and brine, dried with Na₂SO₄, filtered, and concentrated. Theresidue was treated with 0.1 N NaOH (200 mL) and extracted with hexane(2×200 mL). The aqueous layer was brought to pH=2 with 1 N HCl andextracted with EtOAc (2×200 mL). The organic layer was dried with Na₂SO₄and concentrated to give 8 g of acid. ¹H NMR (300 MHz, CDCl₃): d 5.7 (m,1H), 5.36 (d, J=8.4 Hz, 1H), 5.11 (s, 1H), 4.92 (m, 2H), 4.41 (q, J=5.7Hz, 1H), 3.82 (dd, J=9.9, 21.6 Hz, 1H), 3.52 (m, 2H), 3.34 (m, 3H), 2.21(m, 1H), 1.95 (m, 3H), 1.52 (dt, J=7.8, 14.4 Hz, 2H), 1.34 (s, 9H).LC/MS=369.9 (M⁺+1), 392.0 (M⁺+Na)

Step 2. Intermediate IX (6.0 g, 12.9 mmol) was treated with 4 NHCl/1,4-dioxane (32 mL) and stirred for 1 h. The reaction mixture wasconcentrated and dried under vacuum for 20 minutes. The crude amine HClsalt was dissolved in DMF (70 mL) and acid (7.1 g, 25.8 mmol) was added.HATU (12 g, 32.25 mmol) and NMM (6.6 g, 64.5 mmol) were added. Thereaction mixture was stirred at r.t. overnight. The reaction was dilutedwith EtOAc (300 mL), washed with 1 N HCl (200 mL), saturated NaHCO₃,brine, dried with Na₂SO₄, and concentrated. The crude product waspurified by combi-flash to give 7.5 g of dipeptide. ¹H NMR (300 MHz,CDCl₃): d 7.76 (m, 4H), 5.81 (m, 2H), 5.35 (m, 1H), 5.19 (m, 1H), 5.01(m, 2H), 4.73 (m, 1H), 4.48 (m, 1H), 4.03 (m, 1H), 3.79 (m, 1H), 3.68(m, 3H), 3.63 (m, 1H), 3.43 (m, 3H), 2.41 (m, 1H), 2.11 (m, 2H), 1.67(m, 2H), 1.48 (m, 9H).

To a solution of dipeptide (7.1 g, 11.5 mmol) in CH₂Cl₂ (5 mL) was added4 N HCl in 1,4-dioxane (29 mL, 115 mmol). The reaction mixture wasstirred at r.t. for 1 h and concentrated in vacuo for 20 minutes.Cyclopentanol (4.9 g, 57.5 mmol) was dissolved in THF (70 mL) andphosgene (9.7 g, 97.8 mmol) in toluene was added. The reaction wasstirred for 1.5 h and concentrated to half volume to remove phosgene.The crude amine HCl salt was dissolved in EtOAc (200 mL). Na₂CO₃ (17.1g, 138 mmol) in H₂O (100 mL) was added and stirred for 5 minutes. TheTHF solution of the freshly prepared cyclopentylchloroformate was added.The reaction was completed within 1.5 h. The two layers were separated.The organic layers were washed with brine, dried with Na₂SO₄, filtered,and concentrated. The crude product was purified by combi-flash to give3.9 g of product. ¹H NMR (300 MHz, CDCl₃): d 7.76 (m, 4H), 5.81 (m, 2H),5.35 (m, 1H), 5.19 (m, 1H), 5.01 (m, 2H), 4.73 (m, 1H), 4.48 (m, 1H),4.03 (m, 1H), 3.79 (m, 1H), 3.68 (m, 3H), 3.63 (m, 1H), 3.43 (m, 3H),2.41 (m, 1H), 2.11 (m, 2H), 1.67-1.31 (m, 10H).

Ester obtained above (3.9 g, 6.2 mmol) was dissolved in THF (30 mL), H₂O(30 mL), and MeOH (10 mL) and LiOH (1.3 g, 31 mmol) was added. Thereaction mixture was stirred at r. t. for 1 h and diluted with EtOAc.The reaction mixture was acidified to pH=2 with 1 N HCl and separated.The aqueous layer was extracted with EtOAc. The combined organic layerswere washed with brine, dried with Na₂SO₄, concentrated and dried undervacuum to give acid. The crude acid and amine (Example 147, step 1) (2.1g, 6.8 mmol) were dissolved in DMF (50 mL). HATU (5.9 g, 15.5 mmol) andNMM (3.1 g, 31 mmol) were added and the mixture was stirred at r.t. for2 h. The reaction was diluted with EtOAc and washed with 1 N HCl,saturated NaHCO₃/brine, dried with Na₂SO₄ and concentrated. The crudeproduct was purified by combi-flash to give 3.7 g of desired dienecompound. ¹H NMR (300 MHz, CDCl₃): d 7.76 (m, 4H), 7.19 (m, 1H), 6.82(m, 1H), 5.95-5.62 (m, 3H), 5.49-5.20 (m, 3H), 5.19 (m, 1H), 5.01 (m,2H), 4.73 (m, 1H), 4.48 (m, 1H), 4.19-3.82 (m, 3H), 3.79-3.21 (m, 7H),2.41 (m, 1H), 2.23 (m, 1H), 2.11 (m, 2H), 1.83-1.31 (m, 12H). 1.09 (m,3H). ³¹P NMR (121.4 MHz, CDCl₃):

45.168, 43.313 diastereomers.

Diene (3.7 g, 4.3 mmol) was dissolved in CH₂Cl₂ (400 mL) and degassedwith N₂ for 30 minutes. Grubb's G1 (1.1 g, 1.3 mmol) was added anddegassed for an additional 30 minutes. The reaction mixture was heatedto 45° C. for 3 h and cooled to rt. Tris(hydroxymethyl)phosphine (8.0 g,64.5 mmol) was added followed by addition of TEA (13.1 g, 129 mmol) andH₂O (30 mL). The reaction mixture was stirred overnight and two layerswere separated. The organic layer was washed with 0.5 N HCl, brine,dried with Na₂SO₄, and concentrated. The crude product was purified bycombi-flash to give 1.7 g of cyclized compound. ¹H NMR (300 MHz, CDCl₃):d 7.76 (m, 4H), 7.19 (m, 1H), 6.82 (m, 1H), 5.81 (m, 1H), 5.58 (m, 1H),5.19 (m, 1H), 5.01 (m, 2H), 4.73 (m, 1H), 4.48 (m, 1H), 4.19-3.82 (m,3H), 3.79-3.21 (m, 7H), 2.41 (m, 1H), 2.23 (m, 1H), 2.11 (m, 2H),1.83-1.31 (m, 12H). 1.09 (m, 3H). ³¹P NMR (121.4 MHz, CDCl₃):

45.768, 41.813 diastereomers.

A solution of cyclized compound (1.7 g, 1.9 mmol) and8-chloro-2-(2-isopropylamino-thiazol-4-yl)-7-methoxy-quinolin-4-ol (0.62g, 1.8 mmol) in NMP (40 mL) was treated with Cs₂CO₃ (1.3 g, 3.8 mmol).The reaction mixture was heated to 80° C. for 6 h and then cooled to rt.The reaction was diluted with EtOAc and washed with H₂O. The aqueouslayer was brought to pH=4 and extracted with 5% MeOH/EtOAc (2×200 mL).The combined organic layers were dried with Na₂SO₄ and concentrated. Theresidue was purified by combi-flash to give 1.6 g of product.

Ester obtained above (1.6 g, 1.6 mmol) was dissolved in CH₃CN (3 mL) andcooled to 0° C. Iodotrimethylsilane (1.6 g, 8.1 mmol) was addeddropwise. The reaction mixture was warmed to r.t. and stirred for 0.5 h.The reaction was cooled to 0° C. and 2,6-lutidine (1 mL) was added. MeOH(1 mL) was added and the reaction mixture was concentrated. The crudeproduct was purified by HPLC to give 600 mg of acid, which was reducedwith p-tosyl hydrazide (758 mg, 4.1 mmol) and NaOAc (672 mg, 8.2 mmol)to give 160 (390 mg). ¹H NMR (300 MHz, CD₃OD): d 8.32 (d, J=9.6 Hz, 1H),8.26 (s, 1H), 7.86 (s, 1H), 7.65 (d, J=9.3 Hz, 1H), 7.26 (m, 1H), 6.92(t, J=6.9 Hz, 2H), 5.86 (s, 1H), 4.75 (m, 1H), 4.65 (m, 1H), 4.48 (m,2H), 4.21 (m, 1H), 4.18 (s, 3H), 4.07 (m, 1H), 3.75 (m, 6H), 3.53 (m,2H), 2.83 (m, 1H), 2.58 (m, 1H), 1.87 (m, 3H), 1.68-1.36 (m, 17H), 1.36(m, 6H). ³¹P NMR (121.4 MHz, CD₃OD):

42.637. LC/MS=959 (M⁺+1)

Example 161 Preparation of Compound 161

Step 1. 2-Oxa-5-aza-bicyclo[2.2.1]heptan-3-one amine HCl salt (1.0 g,6.7 mmol) and acid (Example 160, step 1) (2.73 g, 10 mmol) weredissolved in CH₂Cl₂ (60 mL)/DMF (7 mL). HATU (7.64 g, 20.1 mmol) and NMM(2.94 mL, 26.80 mmol) were added and the mixture was stirred at r.t. for1 h. The reaction was diluted with CH₂Cl₂ and washed with 5% LiCl (2×).The organic layer was washed with saturated NaHCO₃, dried with Na₂SO₄,and concentrated. The crude product was purified by combi-flash followedby HPLC to give 725 mg of compound in 29% yield.

Step 2. To a yellow solution of product from step 1 (725 mg, 1.97 mmol)and amine example 58 (1.0 g, 3.30 mmol) in toluene (5 mL) was added asolution of sodium hexanoate (327 mg, 1.97 mmol) in H₂O (15 mL). Thereaction mixture was heated to 70° C. for 60 h. The reaction mixture wasdiluted with EtOAc, washed with saturated NH₄Cl (2×), saturated NaHCO₃(2×) and brine. The organic layer was dried with Na₂SO₄, concentrated,and dried under vacuum to give 520 mg of alcohol as crude product.

Alcohol (520 mg, 0.78 mmol) and DABCO (279 mg, 2.48 mmol) were dissolvedin toluene (4 mL). A toluene (4 mL) solution of brosylchloride (635 mg,2.48 mmol) was added dropwise. The reaction mixture was stirred at r. t.for 1 h. The reaction was diluted with EtOAc and quenched with saturatedNaHCO₃. The two layers were separated and the organic layer was washedwith 0.5 N HCl, brine, dried with Na₂SO₄, filtered, and concentrated.The crude product was purified by combi-flash to give 621 mg ofbrosylate in 90% yield. ³¹P NMR (121.4 MHz, CDCl₃)

47.0, 44.5, 42.8, 41.6. LC/MS=890.4 (M⁺+1)

Step 3. Brosylate (620 mg, 0.70 mmol) was dissolved in CH₂Cl₂ (70 mL)and degassed with N₂ for 20 minutes. Grubb's G1 (144 mg, 0.18 mmol) wasadded and degassed for an additional 20 minutes. The reaction mixturewas heated to 50° C. for 5.5 h and cooled to rt.Tris(hydroxymethyl)phosphine (1.09 g) was added followed by addition ofTEA (2.44 mL) and H₂O (10 mL). The reaction mixture was heated to 50° C.for 6 h and then r.t. overnight. The two layers were separated. Theorganic layer was washed with 0.5 N HCl and brine, dried with Na₂SO₄,and concentrated. The crude product was purified by combi-flash to give504 mg of olefin in 84% yield. ¹H NMR (300 MHz, CDCl₃):

³¹P (121.4 MHz, CDCl₃): d

Step 4. A solution of compound olefin (504 mg, 0.59 mmol) and2-(2-isopropylamino-thiazol-4-yl)-7-methoxy-quinolin-4-ol (185 mg, 0.59mmol) in NMP (5.9 mL) was treated with Cs₂CO₃ (191 mg, 0.59 mmol). Thereaction mixture was heated to 63° C. for 7 h and then cooled to rtovernight. The reaction was diluted with EtOAc and washed with NaHCO₃.The organic layer was washed with 5% LiCl and brine, dried with Na₂SO₄,and concentrated. The crude product (551 mg) was used directly for nextstep reaction.

To a solution of crude product obtained above (551 mg, 0.59 mmol) inCH₃CN (5.9 mL) at 0° C. was added iodotrimethylsilane (0.42 mL, 2.94mmol). The reaction mixture was warmed to r.t, stirred for 30 minutes,and cooled to 0° C. 2,6-lutidine (0.69 mL, 5.87 mmol) was added followedby addition of MeOH (3 mL) and warmed to rt. The mixture wasconcentrated, dried under vacuum, and then dissolved in CH₃CN (3 mL).Saturated Na₂CO₃ in H₂O (3 mL) was added and stirred for 5 minutes. ATHF solution of the freshly prepared cyclopentylchloroformate (4.11mmol) was added. The reaction was completed within 1 h. The solvent wasremoved on rotavap. The residue was worked up and purified by HPLC togive 180 mg of 161. ¹H NMR (300 MHz, CD₃OD)

8.27 (d, J=9.6 Hz, 1H), 8.2 (s, 1H), 7.76 (m, 2H), 7.32 (dd, J=2.4, 9.3Hz, 1H), 7.26 (m, 1H), 6.94 (t, J=7.5 Hz, 2H), 5.84 (s, 1H), 5.67 (dd,J=8.4, 9.9 Hz, 1H), 5.45 (t, J=9.9 Hz, 1H), 4.77 (t, J=8.1 Hz, 1H), 4.69(d, J=12 Hz, 1H), 4.54 (m, 2H), 4.17 (m, 3H), 4.03 (s, 3H), 3.26-3.73(brm, 7H), 2.81 (dd, J=7.8, 14.1 Hz, 1H), 2.5-2.7 (brm, 2H), 2.38 (m,1H), 2.2 (m, 1H), 1.4-1.7 (brm, 10H), 1.36 (d, J=15.6 Hz).

³¹P NMR (121.4 MHz, CD₃OD) d 40.9. LC/MS=925.5 (M⁺+1)

Example 162 Preparation of Compound 162

To a solution of 161 (103 mg, 0.11 mmol) in DME (18 mL)/H₂O (1.8 mL) wasadded p-tosylhydrazide (156 mg, 0.84 mmol) and NaOAc (138 mg, 1.68mmol). The reaction mixture was heated to 95° C. for 3 h and cooled tort. The mixture was concentrated, dissolved in CH₂Cl₂, and washed withH₂O. The aqueous layer was extracted with CH₂Cl₂ (2×). The organic layerwas dried with Na₂SO₄ and concentrated. The crude product was purifiedby HPLC to give 74 mg of acid 162 in 71% yield. ¹H NMR (300 MHz, CD₃OD)

8.25 (d, J=9 Hz, 1H), 8.19 (s, 1H), 7.74 (d, J=3.6 Hz, 2H), 7.34 (dd,J=2.1, 9.3 Hz, 1H), 7.26 (qd, J=2.1, 8.7 Hz, 1H), 6.95 (t, J=7.8 Hz,2H), 5.84 (s, 1H), 4.7 (t, J=8.4 Hz, 1H), 4.61 (brs, 2H), 4.52 (dd,J=3.0, 3.6 Hz, 1H), 4.17 (m, 2H), 4.03 (s, 3H), 3.67 (m, 3H), 3.45 (m,1H), 3.33 (m, 2H), 2.82 (dd, J=7.8, 14.4 Hz, 1H), 2.53 (ddd, J=3.9, 8.7,13.5 Hz, 1H), 1.4-2.0 (brm, 19H), 1.34 (d, J=6.3 Hz). ³¹P NMR (121.4MHz, CD₃OD) d 42.9

LC/MS=927.6 (M⁺+1).

Example 163 Preparation of Compound 163

A solution of 2-allyloxy-ethanol (20 g, 196 mmol) in pyridine (3.48 mL,43 mmol) was added to PBr₃ (7.45 mL, 78.4 mmol) at 0° C. The reactionmixture was stirred overnight at rt. The white precipitate was filtered.The organic was diluted with ether, washed saturated NaHCO₃ and brine,dried with Na₂SO₄, filtered, and concentrated. The crude product waspurified by combi-flash to give 6.3 g of 3-(2-bromo-ethoxy)-propene in85% yield. ¹H NMR (300 MHz, CDCl₃):

5.85 (m 1H) 5.19-5.38 (m, 2H), 4.15 (d, 2H), 3.78 (dd, 2H), 3.48 (dd,2H). LC/MS=165 (M⁺+1), 187 (M⁺+Na)

A solution of N-Boc-L-serine (7 g, 34 mmol) in DMSO (100 mL) was treatedwith 60% of NaH (1.36 g, 34 mmol). The mixture was stirred at rt for 1 hand compound 3-(2-bromo-ethoxy)-propene (5.6 g, 34 mmol) was added. Anadditional NaH (1.36 g, 34 mmol) was added and the reaction mixture wasstirred overnight. The reaction was cooled to 0° C., diluted with EtOAc,and quenched slowly with 1N HCl to pH=4. The two layers were separated.The organic layer was washed with H₂O and brine, dried with Na₂SO₄,filtered, and concentrated. The residue was treated with 0.3 N NaOH andextracted with hexane twice. The aqueous layer was brought to pH=4 with1 N HCl and extracted with EtOAc. The organic layer was dried withNa₂SO₄ and concentrated to give 3.75 g of acid. ¹H NMR (300 MHz, CDCl₃):d 5.90 (m, 1H), 5.19-5.38 (m, 2H), 4.42 (m, 1H), 4.15 (dd, 2H), 3.98(dd, 2H), 3.60 (d, 2H), 3.70 (d, 2H). LC/MS=289.9 (M⁺+1)

To a solution of intermediate XI (4 g, 5.45 mmol) in CH₂Cl₂ (30 mL) wasadded 4 N HCl in 1,4-dioxane (32 mL, 129 mmol). The reaction mixture wasstirred at r.t. for 0.5 h, concentrated, dried under vacuum for 20minutes to give amine HCl salt. The resulting amine HCl salt and acidobtained above (2.4 g, 8.42 mmol) were dissolved in DMF (30 mL). HATU (5g, 13.1 mmol) and NMM (2.8 mL, 26 mmol) were added and the mixture wasstirred at r.t. overnight. The crude product was purified by combi-flashto give 3.65 g of tripeptide in 75% yield. ¹H NMR (300 MHz, CDCl₃):

7.80 (m, 4H), 7.18 (m, 1H), 6.89 (m, 2H), 5.98 (m, 2H), 5.42-5.02 (m,5H), 4.78 (m, 1H), 4.58 (m, 1H), 4.20-3.³¹ (m, 12H), 2.82 (s, 2H),2.58-2.21 (m, 2H), 1.80 (m, 1H), 1.42 (s, 9H), 1.38-1.08 (m, 5H). ³¹P(121.4 MHz, CDCl₃):

44.89, 44.82.

LC/MS=905.80 (M⁺+1), 928.13 (M⁺+Na)

To a solution of tripeptide (3.65 g, 4 mmol) in CH₂Cl₂ (20 mL) was added4 N HCl in 1,4-dioxane (20 mL, 80 mmol). The reaction mixture wasstirred at r.t. for 0.5 h, concentrated, dried under vacuum for 20minutes, and then dissolved in EtOAc (50 mL). Na₂CO₃ (5.9 g, 48 mmol) inH₂O (25 mL) was added and stirred for 5 minutes. A THF solution of thefreshly prepared cyclopentylchloroformate (2.93 g, 19.73 mmol) wasadded. The reaction was completed within 1.5 h. The solvent was removedon rotavap and the residue was diluted with EtOAc. The mixture wasbrought to pH=2 with 1 N HCl and the two layers were separated. Theorganic layers were washed with brine, dried with Na₂SO₄, filtered, andconcentrated. Cyclopentyl carbamate (3.5 g) was obtained. ¹H NMR (300MHz, CDCl₃): ¹H NMR (300 MHz, CDCl₃): d 7.78 (m, 4H), 7.18 (min 1H),6.90 (m, 2H), 6.18-5.85 (m, 2H), 5.58-4.90 (m, 5H), 4.78 (m, 1H), 4.60(m, 1H), 4.20-3.25 (m, 13H), 2.81 (s, 2H), 2.60-2.20 (m, 2H), 1.9-1.3(m, 9H), 1.30-1.05 (m, 5H) ³¹P (121.4 MHz, CDCl₃): d 44.70, 42.66.LC/MS=917.93 (M⁺+1), 940.98 (M⁺+Na)

Cyclopentyl carbamate (3.26 g, 3.56 mmol) was dissolved in CH₂Cl₂ (450mL) and degassed with N₂ for 30 minutes. Grubb's G1 (880 mg, 1.07 mmol)was added and degassed for an additional 30 minutes. The reactionmixture was heated to 45° C. overnight and cooled to rt.Tris(hydroxymethyl)phosphine (6.7 g, 53.4 mmol) was added followed byaddition of TEA (15 mL, 107 mmol) and H₂O (50 mL). The reaction mixturewas heated to reflux overnight. The two layers were separated. Theorganic layer was washed with 0.5 N HCl, brine, dried with Na₂SO₄, andconcentrated. The crude product was purified by combi-flash to give 2.73g of cyclic tripeptide.

A solution of cyclic tripeptide (1.75 g, 1.97 mmol) and intermediate X(622 mg, 1.78 mmol) in NMP (35 mL) was treated with Cs₂CO₃ (1.24 g, 3.82mmol). The reaction mixture was heated to 80° C. for 6 h and then keptat rt overnight. The reaction was diluted with EtOAc and washed withH₂O. The aqueous layer was brought to pH=4 with 1 N HCl and extractedwith 5% MeOH/EtOAc (2×). The combined organic layers were dried withNa₂SO₄ and concentrated. The crude product was purified by combi-flashto give 2.0 g of olefin intermediate.

To a solution of olefin intermediate (487 mg, 0.49 mmol) in CH₃CN (2.5mL) at 0° C. was added iodotrimethylsilane (0.34 mL, 2.4 mmol). Thereaction mixture was warmed to r.t. and stirred for 30 minutes and thencooled to 0° C. 2,6-Lutidine (0.34 mL) and MeOH (2.5 mL) were added andstirred for 10 minutes. The solvent was concentrated and the crudeproduct was purified by HPLC to give 246 mg of acid 163. ¹H NMR (300MHz, CD₃OD): d 8.32 (d, J=11.4 Hz, 2H), 7.86 (s, 1H), 7.67 (d, J=9.3 Hz,1H), 7.28 (m, 1H), 6.96 (dd, J=7.8, 7.8 Hz, 2H), 5.85 (s, 1H), 4.67 (m,1H), 4.56 (m, 2H), 4.47 (bs, 1H), 4.24 (m, 1H), 4.17 (s, 3H), 4.03 (m,1H), 3.99-3.46 (m, 10H), 3.32 (m, 3H), 2.87 (m, 1H), 2.61 (m, 1H),2.04-1.25 (m, 18H). ³¹P (121.4 MHz, CD₃OD):

42.463.

LC/MS=975.30 (M⁺+1)

Example 164 Preparation of Compound 164

A solution of 2.17 g (9.36 mmol) of methyl2-t-butoxycarbonylamino-3-aminopropanoate and 1.56 mL (11.19 mmol) oftriethylamine in CH₂Cl₂ (20 mL) was stirred at 0° C. as a solution of3-butenesulfonyl chloride in CH₂Cl₂ (3 mL) was added by canula. After 30min, the mixture was stirred at rt for 14 h and concentrated. Theviscous residue was dissolved in ethyl acetate (50 mL) and washed with0.3 M aq. HCl. The aqueous fraction was extracted with ethyl acetate (30mL). The organic fractions were washed with water (×1) and brine (×1),dried (MgSO₄), and concentrated. The residue was purified by combiflash(120 g column) using hexanes and ethyl acetate to obtain 1.54 g (49%) ofthe sulfonamide as light yellow solids.

Methyl 3-(3-butenylsulfonamido)-2-t-butoxycarbonylamino-propanoate (1.54g, 4.58 mmol) was dissolved in 4 N HCl in dioxane and the solution wasstirred at rt for 1 h and concentrated. After drying the residue in highvacuum for 20 min, the residue and triethylamine (1.92 mL, 13.78 mmol)were dissolved in H₂O (23 mL) and THF (23 mL) andcyclopentyloxycarbonyloxy-succinamide (1.10 g, 4.83 mmol) was added tothe solution. The solution was stirred at rt for 3.5 h and concentratedto a half volume before dilution with H₂O (30 mL). The product wasextracted with ethyl acetate (50 mL×2) and the extracts were washed withH₂O (50 mL×1) and saturated aq. NH₄Cl (50 mL×1), dried (MgSO₄), andconcentrated to afford 1.52 g (95%).

The methyl ester (1.52 g, 4.36 mmol) and LiOH (522.5 mg, 21.82 mmol)were dissolved in H₂O (20 mL), methanol (20 mL), and THF (20 mL) at 0°C. and the resulting mixture was stirred at rt for 9 h. The solution wasconcentrated to remove THF and methanol, the resulting concentratedsolution was diluted with H₂O (30 mL) and washed with ethyl acetate (30mL×1). After the aqueous fraction was acidified with 6 N HCl (5 mL), theproduct was extracted with ethyl acetate (40 mL×3). The extracts werewashed with brine (40 mL×1), dried (MgSO₄), and concentrated to obtain1.30 g (89%) of the acid.

The dipeptide XII (3.20 g, 4.36 mmol) was dissolved in 16.5 mL of 4 NHCl in dioxane and stirred at rt for 1 h. The resulting solution wasconcentrated and dried in vacuum. A solution of the resulting residue inDMF (7 mL) was added to a solution of the acid (1.30 g, 3.89 mmol), HATU(2.22 g, 5.83 mmol), and N-methylmorpholine (1.5 mL, 13.64 mmol) in DMF(8 mL) at rt. After 2 h, the solution was stirred with 5% aq. LiClsolution (60 mL) for 20 min and the mixture was further diluted with H₂O(50 mL) before extraction with ethyl acetate (100 mL×2). The organicextracts were washed with 1 N HCl (100 mL×1), saturated aq. NaHCO₃ (100mL×1), and brine (100 mL×1), dried (MgSO₄), and concentrated. Theresidue was purified by combiflash chromatography (120 g column) toobtain 1.³¹ g (35.5%) of the tripeptide.

A solution of the tripeptide (1.30 g, 1.37 mmol) in CH₂Cl₂ (260 mL) wasdegassed for 30 min and Grubbs G1 (283 mg, 0.344 mmol) was added to thesolution. The solution was refluxed for 5 h at 45° C. bath andadditional catalyst (112 mg, 0.137 mmol) was added before heating foradditional 4 h. To the solution were added tris(hydroxymethyl)phosphine(2.98 g, 24 mmol), triethylamine (6.7 mL, 48.07 mmol), and H₂O (55 mL).The resulting mixture was stirred at 50° C. bath for 3 h. The mixturewas diluted with H₂O (200 mL) and saturated aq. NH₄Cl (200 mL) and thetwo layers were separated. The aqueous fraction was further extractedwith CH₂Cl₂ (100 mL×2). The organic fractions were washed with H₂O (250mL×1), dried (MgSO₄), and concentrated. The residue was purified bycombiflash chromatography to obtain 456 mg (36%) of the major product.

The obtained product (456 mg, 0.495 mmol) was dissolved in NMP (5 mL)and 8-chloro-2-(2-isopropylaminothiazol-4-yl)-7-methoxy-quinolin-4-ol(173 mg, 0.495 mmol) and Cs₂CO₃ (323 mg, 0.991 mmol) were added to thesolution. The resulting mixture was stirred at 70° C. bath for 16 h anddiluted with ethyl acetate (25 mL) before stirring with 5% aq. LiClsolution (20 mL) for 30 min. After the two phases were separated, theaqueous fraction was extracted with ethyl acetate (30 mL×1). The organicfractions were washed with H₂O (×1), dried (MgSO₄), and concentrated.The residue was purified by combiflash chromatography using CH₂Cl₂ andmethanol to obtain 259 mg of the product with some impurities.

A mixture of the impure product (286 mg, 0.250 mmol), tosylhydrazide(350 mg, 1.88 mmol), and sodium acetate (308 mg, 3.75 mmol) in DME (2.5mL) and H₂O (0.25 mL) was refluxed at 95° C. bath for 3 h. Additionaltosylhydrazide (350 mg, 1.88 mmol), and sodium acetate (308 mg, 3.75mmol) were added to the mixture and the mixture was refluxed for 1.5 h.The mixture was diluted with H₂O (50 mL) and saturated aq. NaHCO₃ (50mL), and extracted with ethyl acetate (40 mL×2). The extracts werewashed with H₂O (×1), dried (MgSO₄), and concentrated. The residue waspurified by reverse-phase combiflash chromatography (43 g column) usingH₂O and acetonitrile with 0.05% TFA. The combined fractions wereconcentrated to remove acetonitrile, and the product was extracted withethyl acetate (×2) from the resulting aq. solution was diluted withsaturated aq. saturated NaHCO₃ to obtain 135 mg of the product with someimpurities.

A solution of the impure product (135 mg, 0.130 mmol) and 2,6-lutidine(0.19 mL, 1.63 mmol) in acetonitrile (3 mL) was stirred at 0° C. as TMSI(0.18 mL, 1.27 mmol) was added. After the mixture was stirred at rt for2 h, methanol (2 mL) was added and the solution was stirred for 1 h atrt. The solution was concentrated and the residue was purified byreverse-phase HPLC. The product containing fractions were pooled,concentrated and freeze-dried to obtain 74.6 mg (51%) as TFA salt. ^(1H)NMR (300 MHz, CD₃OD): d 8.26 (d, 1H, J=9.3 Hz), 8.24 (s, 1H), 7.78 (s,1H), 7.60 (s, 1H), 7.24 (appt quint, 1H, J=7.2 Hz), 6.92 (t, 1H, J=7.5Hz), 5.83 (br, 1H), 4.67-4.78 (m, 2H), 4.74 (dd, 1H), 4.40 (br, 1H),4.21 (d, 1H, J=12.0 Hz), 4.15 (s, 3H), 3.99 (hept, 1H, J=6.3 Hz),3.07-3.54 (m, 6H), 2.89 (dd, 1H, J=12.9 and 6.9 Hz), 2.44-2.59 (m, 1H),1.92-2.26 (m, 3H), 1.15-1.81 (m, 16H), 1.38 (d, 6H, J=6.3 Hz). ³¹P(121.4 MHz, CD₃OD): d 40.177. ¹⁹F (282 MHz, CD₃OD): d −77.³¹1 (CF₃COOHsalt), −114.754. LC/MS=1008 (M⁺+1)

Example 165 Preparation of Compound 165

Step 1. To a yellow solution of VIII (477 mg, 1.30 mmol) and(1-amino-2-vinyl-cyclopropyl)-(2-methoxy-benzyl)-phosphinic acid ethylester (Example 42) (274 mg, 0.93 mmol) in toluene (4.5 mL) was added asolution of sodium 2-ethyl-hexanoate (77 mg, 0.46 mmol) in H₂O (4.5 mL).The reaction mixture was heated to 70° C. for 22 h. Additional2-ethyl-hexanoate (100 mg) was added, stirred for 22 h at 70° C. andcooled to rt. The reaction mixture was diluted with EtOAc, washed withsaturated NH₄Cl (2×), saturated NaHCO₃ (2×) and brine. The organic layerwas dried with Na₂SO₄ and concentrated. The crude product was purifiedby column chromatography to give 468 mg of alcohol. LC/MS=661.9 (M⁺+1),683.9 (M⁺+Na)

Step 2. Alcohol (468 mg, 0.71 mmol) and DABCO (255 mg, 2.27 mmol) weredissolved in toluene (3.5 mL). A toluene (3.5 mL) solution of brosylchloride (580 mg, 2.27 mmol) was added dropwise. The reaction mixturewas stirred at r. t. for 1 h. The reaction was diluted with EtOAc andquenched with saturated NaHCO₃. The two layers were separated and theorganic layer was washed with 0.5 N HCl, brine, dried with Na₂SO₄,filtered, and concentrated. The crude product was purified bycombi-flash to give 402 mg of brosylate in 49% yield. ³¹P NMR (121.4MHz, CDCl₃)

46.5, 44.1. LC/MS=881.4 (M⁺+1), 903.8 (M⁺+Na)

Step 3. To a solution of brosylate (402 mg, 0.46 mmol) in CH₂Cl₂ (3.4mL) was added 4 N HCl in 1,4-dioxane (3.4 mL, 17 mmol). The reactionmixture was stirred at r.t. for 2 h, concentrated, dried under vacuumovernight, and then dissolved in THF (3.4 mL). The freshly preparedcyclopentylchloroformate (2.33 mmol) in THF (4.6 mL) was added. TEA(0.32 mL, 2.28 mmol) was added to the reaction mixture. The reaction wascompleted within 1 h. The reaction was quenched by adding saturatedNH₄Cl and diluted with EtOAc. The two layers were separated. The organiclayers were washed with saturated NH₄Cl and brine, dried with Na₂SO₄,and concentrated. The crude product was purified by columnchromatography to give 349 mg of cyclopentyl carbamate in 86% yield. ³¹PNMR (121.4 MHz, CDCl₃)

46.4, 44.1. LC/MS=893.8 (M⁺+1), 915.6 (M⁺+Na)

Step 4. Cyclopentyl carbamate (349 mg, 0.39 mmol) was dissolved inCH₂Cl₂ (28 mL) and degassed with N₂ for 20 minutes. Grubb's G1 (80 mg,0.10 mmol) was added and degassed for an additional 20 minutes. Thereaction mixture was heated to 50° C. for 6.5 h. Additional Grubb's G1(40 mg) was added and heated to 60° C. for 8 h. More Grubb's G1 (40 mg)was added, stirred at 50° C. for 5 h, and cooled to rt.Tris(hydroxymethyl)phosphine (1.21 g) was added followed by addition ofTEA (2.7 mL) and H₂O (5 mL). The reaction mixture was heated to 50° C.for 6 h and then r.t. overnight. The two layers were separated. Theorganic layer was washed with 0.5 N HCl and brine, dried with Na₂SO₄,and concentrated. The crude product was purified by combi-flash to give210 mg of compound olefin in 62% yield.

LC/MS=865.5 (M⁺+1), 887.5 (M⁺+Na)

Step 5. A solution of compound olefin (210 mg, 0.24 mmol) and2-(2-isopropylamino-thiazol-4-yl)-7-methoxy-quinolin-4-ol (77 mg, 0.24mmol) in NMP (2.4 mL) was treated with Cs₂CO₃ (79 mg, 0.24 mmol). Thereaction mixture was heated to 65° C. for 6 h and then cooled to rtovernight. The reaction was diluted with EtOAc and washed with NaHCO₃.The organic layer was washed with 5% LiCl and brine, dried with Na₂SO₄,and concentrated. The crude product (229 mg, 0.24 mmol) was dissolved inCH₃CN (3 mL), 2,6-lutidine was added at 0° C. was added, followed byiodotrimethylsilane (0.52 mL, 3.65 mmol). The reaction mixture waswarmed to rt, stirred for 3 h, and cooled to 0° C. 2,6-lutidine (0.2 mL)was added followed by addition of MeOH (2.5 mL) and warmed to rt. Themixture was concentrated and purified by HPLC to give 98 mg of product165 in 44% yield. ¹H NMR (300 MHz, CD₃OD)

8.13 (d, J=9.3 Hz, 1H), 7.99 (s, 1H), 7.55 (s, 2H), 7.09 (m, 2H), 7.00(m, 1H), 6.73 (d, J=8.4 Hz, 1H), 6.67 (t, J=7.5 Hz, 1H), 5.63 (s, 1H),5.45 (q, J=9 Hz, 1H), 5.08 (t, i=9.9 Hz, 1H), 4.7 (d, J=12 Hz, 1H), 4.55(t, J=9.3 Hz, 1H), 4.24 (s, 1H), 3.99 (m, 2H), 3.89 (dd, J=2.4, 11.7 Hz,1H), 3.83 (s, 3H), 3.62 (s, 3H), 3.29 (t, J=15 Hz, 1H), 3.10 (t, J=15Hz, 1H), 2.6 (dd, J=7.5, 14.1 Hz, 1H), 2.6 (m, 1H), 2.42 (ddd, J=3.6,9.6, 13.5 Hz, 1H), 2.05 (m, 1H), 1.64 (m, 2H), 1.22-1.46 (brm, 17H),1.16 (d, J=6.5 Hz, 6H). ³¹P NMR (121.4 MHz, CD₃OD)

43.4

LC/MS=914.6 (M⁺+1)

Example 166 Preparation of Compound 166

To a solution of 165 (887 mg, 0.97 mmol) in DME (87 mL)/H₂O (8.7 mL) wasadded p-tosyl hydrazide (1.45 g, 7.28 mmol) and NaOAc (1.19 g, 14.57mmol). The reaction mixture was heated to 95° C. for 4 h and cooled tort. The mixture was concentrated, dissolved in CH₂Cl₂, and washed withH₂O. The aqueous layer was extracted with CH₂Cl₂ (2×). The organic layerwas dried with Na₂SO₄, concentrated, and dried under vacuum for 30minutes. The residue was dissolved in DME (60 mL) and H₂O (6 mL). NaOAc(1.19 g, 14.57 mmol) and p-tosylhydrazide (1.45 g, 7.28 mmol) wereadded. The reaction mixture was heated to 95° C. for 6 h and cooled tort. The reaction was worked up and purified by HPLC to give 318 mg ofacid 166 in 36% yield, ¹H NMR (300 MHz, CD₃OD)

8.23 (d, J=9.6 Hz, 1H, 8.18 (s, 1H), 7.69 (s, 1H), 7.31 (dt, J=1.8, 7.2Hz, 1H), 7.25 (dd, J=2.1, 9.3 Hz, 1H), 7.18 (t, J=7.8 Hz, 1H), 6.92 (d,J=8.1 Hz, 1H), 6.87 (t, J=7.5 Hz, 1H), 5.79 (s, 1H), 4.8 (d, J=11.7 Hz,1H), 4.69 (t, J=8.7 Hz, 1H), 4.47 (s, 1H), 4.23 (dd, J=2.7, 10.2 Hz,1H), 4.15 (t, J=6.3 Hz, 1H), 4.1 (d, J=11.1 Hz, 1H), 3.98 (s, 3H), 3.81(s, 3H), 3.43 (t, J=15 Hz, 1H), 3.32 (t, J=15 Hz, 1H), 2.76 (dd, J=8.1,14.4 Hz, 1H), 2.49 (ddd, J=3.6, 9, 13.2 Hz, 1H), 1.98 (m, 1H), 1.79 (m,2H), 1.34 (d, J=6.3 Hz, 1H), 1.2-1.7 (m, 22H). ³¹P NMR (121.4 MHz,CD₃OD)

45.5. LC/MS=915.3 (M⁺+1).

Example 167 Preparation of Compound 167

The product (Example 167) was afforded as a yellow solid.

¹H NMR (300 MHz, CD₃OD):

8.32 (d, J=9.3 Hz, 1H), 8.18 (s, 1H), 7.71 (m, 2H), 7.67 (m, 2H), 7.53(m, 1H), 7.37 (m, 2H), 5.86 (s, 1H), 5.75 (m, 1H), 5.34 (m, 1H), 4.73(m, 1H), 4.11 (m, 3H), 4.04 (s, 3H) 3.72 (m, 1H), 3.38 (m, 1H), 2.80 (m,3H), 2.64 (m, 1H), 2.31 (m, 1H) 1.80 (m, 1H), 1.65-1.2 (m, 24H

³¹P NMR (121.4 MHz, CD₃OD): d 39.599. LC (6 minute run, r.t.=3.67 min)MS (954.6, M+1)

Example 168 Preparation of Compound 168

The product (Example 168) was obtained as a yellow solid. ¹H NMR (300MHz, CD₃OD):

8.29 (d, J=9.5 Hz, 1H), 8.17 (s, 1H), 7.69 (m, 4H), 7.56 (m, 1H), 7.37(m, 2H), 5.86 (s, 1H), 4.80 (m, 1H), 4.74 (m, 1H) 4.38 (s, 1H) 4.11 (m,3H), 4.04 (s, 3H) 3.30-3.62 (m, 3H), 2.80 (m, 1H), 2.59 (m, 1H), 2.01(m, 1H) 1.78 (m, 1H), 1.65-1.2 (m, 26H). ³¹P NMR (121.4 MHz, CD₃OD):

418

LC (6 minute run, r.t.=3.65 min) MS (956.5, M+1).

Example 169 Preparation of Compound 169

To a solution of phosphinate (described in Example 51) (5.0 g, 11.55mmol) in TFA (31 mL, 416 mmol) at r.t. was added DMS (8.2 mL, 111 mmol)and stirred overnight. The reaction mixture was poured into ice cold 4 NHCl (350 mL) and extracted with 1/1 iPrOH/heptane (420 mL). The organiclayers were washed with 4 N HCl (5×500 mL). The combined aqueous layerswere brought to pH=10 in a cold bath. The aqueous layer was extractedwith EtOAc (5×500 mL). The organic layers were washed with brine, driedwith Na₂SO₄, and concentrated to give 2.9 g of amine in 84% yield. ¹HNMR (300 MHz, CDCl₃): 7.58 (m, 1H), 7.41 (m, 1H), 7.18 (m, 2H), 5.96 (m,1H), 5.60 (m, 1H), 5.19 (m, 2H), 5.09 (m, 1H), 4.03 (m, 2H), 3.44 (m,2H), 1.83 (m, 1H), 1.32 (m, 3H), 1.19 (m, 2H). ³¹P NMR (121.4 MHz,CDCl₃):

49.684, 47.512

To a yellow solution of VIII (3.5 g, 9.56 mmol) and amine (2.2 g, 7.36mmol) in toluene (20 mL) was added a solution of sodium hexanoate (1.83g, 11.04 mmol) in H₂O (60 mL). The reaction mixture was heated to 80° C.for 40 h and cooled to rt. The reaction mixture was diluted with EtOAc,washed with saturated NaHCO₃, 0.5 N HCl, brine, dried with Na₂SO₄, andconcentrated. The crude product was purified by combi-flash to give 4.4g of alcohol in 69% yield. ¹H NMR (300 MHz, CDCl₃

d 7.48 (m, 1H), 7.36 (m, 1H), 7.18 (m, 2H), 6.07 (m, 1H), 5.78 (m, 2H),5.50 (m, 1H), 5.30 (m, 1H), 5.17 (m, 1H), 4.67 (d, 1H), 4.48 (m, 2H),4.33 (m, 2H), 4.12 (m, 2H), 3.83 (m, 2H), 3.65 (m, 2H), 3.32 (m, 3H),2.24 (m, 3H), 2.04 (m, 4H), 1.80 (m, 2H), 1.63 (m, 1H), 1.51 (m, 2H),1.42 (m, 9H), 1.25 (m, 3H). ³¹P NMR (121.4 MHz, CDCl₃):

45.191, 43.161 diastereomers.

Alcohol (1.4 g, 2.1 mmol) and DABCO (750 mg, 6.7 mmol) were dissolved intoluene (20 mL). A toluene solution of brosylchloride (1.7 g, 6.7 mmol)was added dropwise. The reaction mixture was stirred at r. t. for 3 h.The reaction was diluted with EtOAc and quenched with saturated NaHCO₃.The two layers were separated and the organic layer was washed with 0.5N HCl, brine, dried with Na₂SO₄, filtered, and concentrated. The crudeproduct was purified by combi-flash to give 1.3 g of brosylate in 70%yield. ¹H NMR (300 MHz, CDCl₃

d 7.74 (m, 4H), 7.48 (m, 1H), 7.36 (m, 1H), 7.18 (m, 2H), 6.07 (m, 1H),5.78 (m, 2H), 5.25 (m, 1H), 5.17 (m, 1H), 4.98 (m, 1H), 4.57 (d, 1H),4.48 (m, 2H), 4.33 (m, 2H), 4.12 (m, 2H), 3.83 (m, 2H), 3.65 (m, 2H),3.32 (m, 3H), 2.24 (m, 3H), 2.04 (m, 4H), 1.80 (m, 2H), 1.63 (m, 1H),1.51 (m, 2H), 1.42 (m, 9H), 1.25 (m, 3H). ³¹P NMR (121.4 MHz, CDCl₃):

46.216, 43.654 diastereomers.

Brosylate (1.48 g, 1.61 mmol) was dissolved in CH₂Cl₂ (200 mL) anddegassed with N₂ for 20 minutes. Grubb's G1 (413 mg, 0.5 mmol) was addedand degassed for an additional 20 minutes. The reaction mixture washeated to 45° C. overnight and cooled to rt.Tris(hydroxymethyl)phosphine (3.4 g, 27.4 mmol) was added followed byaddition of TEA (8 mL, 50 mmol) and H₂O (20 mL). The reaction mixturewas heated to 50° C. for 4 h and then r.t. overnight. The two layerswere separated. The organic layer was washed with 0.5 N HCl, brine,dried with Na₂SO₄, and concentrated. The crude product was purified bycombi-flash to give 907 mg of cyclic phosphinate. ¹H NMR (300 MHz, CDCl₃

d 7.76 (m, 4H), 7.40 (m, 1H), 7.30 (m, 1H), 7.12 (m, 2H), 5.78 (m, 1H),5.60 (m, 1H), 5.40 (m, 1H), 5.07 (m, 1H), 4.90 (m, 1H), 4.37 (m, 3H),3.83 (m, 2H), 3.75 (m, 2H), 3.51 (m, 1H) 3.22 (m, 2H), 2.24 (m, 3H),2.04 (m, 4H), 1.80 (m, 2H), 1.63 (m, 2H), 1.51 (m, 2H), 1.42 (m, 9H),1.25 (m, 3H). ³¹P NMR (121.4 MHz, CDCl₃):

45.216, 44.153 diastereomers.

A solution of cyclic phosphinate (857 mg, 1.0 mmol) and phenol (³¹6 mg,1.0 mmol) in NMP (10 mL) was treated with Cs₂CO₃ (651.6 mg, 2.0 mmol).The reaction mixture was heated to 65° C. overnight and then cooled tort. The reaction was diluted with EtOAc and washed with H₂O. The aqueouslayer was brought to pH=4 with 1 N HCl and extracted with 5% MeOH/EtOAc(2×50 mL). The combined organic layers were dried with Na₂SO₄ andconcentrated. The crude product was purified by combi-flash to give 670mg of desired product. ¹H NMR (300 MHz, CDCl₃): ¹H NMR (300 MHz, CDCl₃):d 8.02 (d, J=9.1 Hz, 1H), 7.47 (s, 1H), 7.37 (m, 2H), 7.15 (m, 2H), 7.02(m, 2H), 5.69 (m, 1H), 5.40 (m, 1H), 5.35 (m, 1H), 5.28 (m, 1H), 4.82(m, 1H), 4.69 (min, 2H), 4.39 (m, 1H), 4.18 (m, 4H), 3.98 (s, 3H), 3.90(m, 2H), 3.80 (m, 2H), 3.42 (m, 1H), 3.23 (m, 1H), 2.71 (m, 1H), 2.40(m, 3H) 2.02 (m, 2H), 1.76 (m, 2H), 1.36 (m, 9H), 1.30 (m, 6H), 1.15 (m,3H). ³¹P NMR (121.4 MHz, CDCl₃):

45.152, 44.291 diastereomers.

To a solution of product obtained above (670 mg, 0.72 mmol) in CH₃CN (2mL) at 0° C. was added iodotrimethylsilane (717 mg, 3.58 mmol). Thereaction mixture was stirred at 0° C. for 20 min. 2,6-Lutidine (1 mL)and MeOH (1 mL) were added, stirred for 20 min, concentrated in vacuo,and dried for 20 minutes to give crude acid. Cyclopentanol (308 mg, 3.58mmol) was dissolved in THF (7.8 mL) and phosgene (3.2 mL, 6.09 mmol) intoluene was added. The reaction was stirred for 1 h and concentrated tohalf volume to remove phosgene. The crude acid was dissolved in CH₃CN(20 mL). Na₂CO₃ (1.1 g, 8.64 mmol) in H₂O (10 mL) was added and stirredfor 5 minutes. The THF solution of the freshly preparedcyclopentylchloroformate was added. The reaction was completed within 1h and concentrated. The residue was dissolved in EtOAc and 1.0 N HCl wasadded to adjust pH=2. The two layers were separated and the organiclayer was concentrated. The crude product was purified by HPLC to giveproduct 169. ¹H NMR (300 MHz, CD₃OD): d 8.89 (s, 1H), 8.34 (d, J=9.5 Hz,2H), 8.18 (s, 1H), 7.76 (m, 2H), 7.39 (m, 1H), 7.30 (min, 1H), 7.21 (m,2H), 5.83 (s, 1H), 5.70 (m, 1H), 5.31 (m, 1H), 4.73 (m, 1H), 4.40 (m,1H), 4.16 (m, 2H), 4.02 (m, 4H), 3.72 (m, 1H), 3.38 (m, 2H), 2.81 (m,1H), 2.62 (m, 1H), 2.29 (m, 1H) 1.95-1.37 (m, 2H), 1.34 (m, 6H). ³¹P NMR(121.4 MHz, CD₃OD):

41.998

LC/MS=919 (M⁺+1)

Example 170 Preparation of Compound 170

To a solution of 169 (50 mg, 0.05 mmol) in DME (1 mL)/H₂O (0.1 mL) wasadded p-tosyl hydrazide (76 mg, 0.41 mmol) and NaOAc (66 mg, 0.81 mmol).The reaction mixture was heated to 95° C. for 1.5 h and cooled to rt. Afew drops of 3 N HCl was added to adjust pH=2. The crude product waspurified by HPLC to give 25 mg of acid 170. ¹H NMR (300 MHz, CD₃OD): d8.89 (s, 1H), 8.30 (d, J=9.4 Hz, 2H), 8.16 (s, 2H), 7.75 (m, 3H), 7.41(m, 4H), 7.22 (m, 3H), 5.84 (s, 1H), 4.74 (m, 2H), 4.42 (m, 1H), 4.18(m, 2H), 4.07 (m, 4H), 3.52 (m, 1H), 3.31 (m, 2H), 2.77 (m, 1H), 2.52(m, 1H), 2.00 (m, 1H), 1.81 (m, 1H), 1.58 (m, 2H), 1.35 (m, 6H). ³¹P NMR(121.4 MHz, CD₃OD)

44.041

LC/MS=921 (M⁺+1)

Example 171 Preparation of Compound 171

Step 1. A solution of the 4-Hydroxy-pyrrolidine-1,2-dicarboxylic acid1-tert-butyl ester (460 mg, 1.99 mmol) and Et₃N (417 μl, 2.99 mmol) inTHF (13 mL) was stirred at −43° C. as ethyl chloroformate (209 μl, 2.19mmol) was added. The mixture was stirred between −40° C. and −25° C. for30 minutes. To this mixture a solution of(1-Amino-2-vinyl-cyclopropyl)-phosphonic acid diethyl ester (480 mg,2.19 mmol) in THF (13 mL) was added drop-wise while the internaltemperature was kept below −35° C. The solution was allowed to warm tort and quenched with H₂O. Reaction mixture was extracted with EtOAc.Combined organic washes were then extracted with brine, 1N HCl (2×) andthen brine. Organic layer was dried over MgSO₄. The crude product (760mg, 88%) was used without further purification. LC/MS=433 (M⁺+1), 454.9(M⁺+Na)

Step 2. To a solution of ester (760 mg, 1.75 mmol) in CH₂Cl₂ (4.40 mL)was added 4N HCl/Dioxane (4.40 mL, 17.5 mmol). The reaction was stirredat rt for 2 h. The reaction was concentrated and the residue was driedunder vacuum for 2 h. The crude amine was used without furtherpurification.

LC/MS=333.3 (M⁺ free base+1), 454.9 (M⁺ free base+Na)

Step 3. To a solution of amine (644 mg, 1.75 mmol),2-tert-Butoxycarbonylamino-non-8-enoic acid and HATU (665 mg, 1.75 mmol)in CH₂Cl₂ (13.125 mL) and DMF (4.375 mL) was added N-methyl-morpholine(524 μL, 4.77 mmol). The reaction was stirred at rt for 30 min. Thereaction was quenched with 5% LiCl (20 mL), and the mixture was stirreduntil the two layers were separated. After the mixture was extractedwith EtOAc (75 mL) the aqueous layer was further extracted with EtOAc.The combined organic layers were washed with saturated NaHCO₃, brine,and then dried over MgSO₄. The filtrate was concentrated and the residuewas purified using column chromatography to obtain tripeptide (440 mg,43%). ³¹P NMR (121.4 MHz, CDCl₃):

22.3. LC/MS=586.0 (M⁺+1), 607.9 (M⁺+Na).

Step 4. A solution of tripeptide (2.4 g, 4.10 mmol) in CH₂Cl₂, which hadbeen degassed by bubbling N₂ through it for 20 min, and Grubb's G1catalyst (844 mg, 1.025 mmol) was heated to 40° C. After the reactionhad stirred at 40° C. for 4 h, additional Grubbs G1 catalyst (150 mg,0.18 mmol) was added and the solution continued to stir at ° C.overnight. Additional Grubbs G1 catalyst (132 mg, 0.16 mmol) was addedand the solution was refluxed in a 50° C. oil bath. After 4 h,tris-(hydroxymethyl)phosphine (8.48 g, 68.34 mmol) was added and thesolution was stirred for 10 minutes before Et₃N (19 mL, 136 mmol) wasadded, followed by H₂O (20 mL). After the mixture was stirred under N₂at 50° C. for 2 h, another portion of H₂O (10 mL) was added and themixture was heated an additional 4 h at 50° C. and then stirredovernight at rt. The aqueous phase was removed and the organic phase waswashed with H₂O, 0.5 N HCl, and saturated NaHCO₃. The organic phase wasthen dried over MgSO₄ and concentrated. The residue was purified usingcolumn chromatography to yield olefin (1.85 g, 81%).

³¹P NMR (121.4 MHz, CDCl₃):

22.160. LC/MS=558.0 (M⁺+1), 579.8 (M⁺+Na).

Step 5. A solution of olefin (1.85 g, 3.32 mmol) and sodium iodide (4.98g, 33.2 mmol) in pyridine (33.2 mL) was heated at 105° C. for 10 h. Thereaction was then at rt for 2 days. Additional sodium iodide (1.50 g,10.0 mmol) was added and the solution was heated at 105° C. for 2 h. Thereaction was then cooled to rt.

To this solution was added 4-Di(methylamino)pyridine (41 mg, 0.33 mmol)and acetic anhydride (4.10 mL, 43.16 mmol). The reaction was stirred atrt for 2 h, and then frozen overnight. The solution was defrosted andEtOAc was added. Saturated NaHCO₃ was added to adjust the pH to 7-8, andthen the layers were separated. The organic layer was extracted with H₂Oand the combined aqueous layers were acidified to pH 1-2 with 1N HCl.The aqueous layers were extracted with 200 mL of ethyl acetate (4×) andthe combined organic layers were washed with brine, dried over MgSO₄,and concentrated. The residue was purified using reverse phase HPLC togive monoacid (1.15 g, 61%) as a white solid. ³¹P NMR (121.4 MHz,CDCl₃):

21.3, 20.9. LC/MS=571.8 (M⁺+1).

Step 6. To a solution of oxalyl chloride (589 μl, 6.75 mmol) in toluene(16.9 mL), dimethylformamide (26 μl, 0.34 mmol) was added drop-wise. Theresulting solution was stirred at rt for 10 minutes. The monoacid (965mg, 1.69 mmol) in toluene (8 mL) was added drop-wise to the previoussolution and the resulting mixture was stirred for 30 minutes at rt. Themixture was concentrated and placed under high vacuum for 20 minutes.The residue was dissolved in THF (16.9 mL) and cooled to −35° C.n-Propylmagnesium chloride (845 μl, 1.69 mmol) was added drop-wise andthe reaction stirred at −30° C. for 30 minutes. Additionaln-propylmagnesium chloride (845 μl, 1.69 mmol) was added and thereaction was warmed to −25° C. and stirred for 20 minutes. Additionaln-propylmagnesium chloride (845 μl, 1.69 mmol) was added and thereaction was stirred at −25° C. for 20 minutes. More n-propylmagnesiumchloride (845 μl, 1.69 mmol) was added and stirred an addition 15minutes at −25° C. A final addition of n-propylmagnesium chloride (845μl, 1.69 mmol) was conducted and the solution stirred at −30° C.Saturated NH₄Cl was used to quench the reaction at −30° C. The reactionwas warmed to rt and diluted with EtOAc. The organic layer was washedwith saturated NH₄Cl and brine, dried over MgSO₄, and concentrated.Desired product (468 mg, 46%) was isolated from the residue by columnchromatography, as white foam. LC/MS=598.0 (M⁺+1), 619.9 (M⁺+Na).

Step 7. To a solution of product obtained above in CH₂Cl₂ (9.8 mL) wasadded 4N HCl/Dioxane (4.90 mL, 19.58 mmol) and stirred for 1.5 h at rt.The solution was concentrated and dried under high vacuum. The whitesolid was used without further purification. LC/MS=497.8 (M⁺ freebase+1), 519.9 (M⁺ free base+Na).

Step 8. A solution of amine in CH₃CN (7.83 mL), H₂O (783 μl) and Et₃N(273 mL, 1.96 mmol) stirred for 5 minutes before cyclopentylchloroformate (173 μl, 1.17 mmol) was added drop-wise. The reactionstirred at rt for 35 minutes and was concentrated and taken up in ethylacetate. The organic phase was washed with saturated NH₄Cl and brine.The organic layer was dried over MgSO₄ and concentrated. Cyclopentylcarbamate was purified from the residue by column chromatography (388mg, 86% over 2 steps).

³¹P NMR (121.4 MHz, CDCl₃):

50.2, 49.6, 48.8, 47.9. LC/MS=609.8 (M⁺+1), 6³¹.0 (M⁺+Na).

Step 9. A solution of cyclopentyl carbamate (388 mg, 0.64 mmol) in THF(9.54 mL) and MeOH (6.36 mL) was stirred at 0° C. as a solution of LiOH(27 mg, 0.64 mmol) in H₂O (3.16 mL) was added drop-wise. The reactionwas then stirred for 45 minutes. The reaction was quenched withsaturated NH₄Cl and extracted with CH₂Cl₂. The organic layer was driedover MgSO₄ and concentrated. The resulting alcohol was placed on highvacuum and used without further purification.

Step 10. To a solution of the alcohol (361 mg, 0.64 mmol) and DABCO (200mg, 1.78 mmol) in toluene (1.27 mL) was added 4-bromobenzenesulfonylchloride (455 mg, 1.78 mmol) in toluene (1.27 mL). The reaction wasstirred at rt for 2 h. The reaction was diluted with toluene andquenched with 1M Na₂CO₃. The organic layer was then extracted with 1MNa₂CO₃, diluted with a small volume of THF and washed with 0.5M HCl. Theorganic layer was then washed with H₂O, dried over MgSO₄ andconcentrated. Brosylate was purified from the residue by columnchromatography (323 mg, 65%) as a white foam/semi-crystalline solid.LC/MS=685.5 (M⁺+1).

Step 11. A solution of brosylate (100 mg, 0.13 mmol),2-(2-Isopropylamino-thiazol-4-yl)-7-methoxy-quinolin-4-ol (40 mg, 0.13mmol) and Cs₂CO₃ (42 mg, 0.13 mmol) in 1-methyl-2-pyrrolidinone (2 mL)was stirred at 60° C. for 10 h, and then overnight at rt. The reactionwas then re-heated to 60° C. and stirred for another 5 h. The reactionwas diluted with EtOAc and washed with a 1:1 solution of H₂O andsaturated NaHCO₃ and then brine. The organic layer was dried over Na₂SO₄and concentrated. The residue was placed on high vacuum overnight. Thecrude (110 mg, 0.03 mmol) and 2,6-lutidine (148 μl, 1.27 mmol) in CH₃CN(2 mL) was stirred at 0° C. as iodotrimethylsilane (181 μl, 1.27 mmol)was added drop-wise. The reaction was stirred at rt for 3.5 h. Thereaction was then cooled to 0° C. and additional 2,6-lutidine (74 μl,0.64 mmol) and iodotrimethylsilane (91 μl, 0.64 mmol) was added. Thereaction was warmed to rt for 1 h. The reaction was cooled to 0° C. andthen Et₃N (200 μl) and MeOH (2.5 mL) was added. The reaction wasconcentrated and 171 (47.6 mg, 45%) was isolated from the residue byHPLC as a yellow solid. ¹H NMR (300 MHz, CD₃OD)

8.30 (d, J=9 Hz, 1H), 8.18 (s, 1H), 7.72 (s, 2H), 7.27 (dd, J=2.4, 9.3Hz, 1H), 5.79 (s, 1H), 5.60 (dd, J=8.1, 17.7 Hz, 1H), 5.21 (t, J=9.6 Hz,1H), 4.58 (s, 1H), 4.69 (t, J=9.3 Hz, 1H), 4.39 (s, 1H), 4.04-4.22 (brm,3H), 4.01 (s, 3H), 2.8 (m, 1H), 2.77 (dd, J=7.2, 14.4 Hz, 1H), 2.56(ddd, J=4.2, 9.9, 14.1 Hz, 1H), 2.20 (m, 1H), 1.2-1.9 (brm, 23H), 1.34(d, J=6.3 Hz, 6H), 1.01 (t, J=7.5 Hz, 3H). ³¹P NMR (121.4 MHz, CD₃OD) d48.2. LC/MS=836.8 (M⁺+1).

Example 172 Preparation of Compound 172

A reaction vessel containing a solution of brosylate (40 mg, 0.05 mmol)and 5% Rh/Al₂O₃ (12 mg) in EtOAc (2 mL) was evacuated under vacuum andthe atmosphere was replaced with H₂. This evacuation/re-pressurizationcycle was repeated 2 more times. The reaction was then stirred under H₂at rt for 16 h. The reaction was filtered through a plug of Celite 541and the filtrate was concentrated. Saturated compound (22 mg, 55%) wasisolated from the residue by column chromatography as a white solid. ³¹PNMR (121.4 MHz, CDCl₃) d 54.2, 50.8. LC/MS=787.5 (M⁺+1).

Replacement and deprotection as described for Example 171 gave compound172. ¹H NMR (300 MHz, CD₃OD):

8.28 (d, J=9.3 Hz, 1H), 8.17 (s, 1H), 7.74 (s, 2H), 7.31 (dd, J=2.1, 9Hz, 1H), 5.81 (s, 1H), 4.81 (d, J=12 Hz, 1H), 4.67 (t, J=8.7 Hz, 1H),4.42 (s, 1H), 4.17 (m, 2H), 4.02 (dd, J=2.7, 12.3 Hz, 1H), 4.03 (s, 3H),2.80 (dd, J=7.8, 15 Hz, 1H), 2.49 (ddd, J=3.9, 9.3, 13.8 Hz, 1H),1.25-1.9 (brm, 29H), 1.34 (d, J=6.3 Hz, 6H), 1.06 (t, J=7.5 Hz, 3H).

³¹P NMR (121.4 MHz, CD₃OD) d 50.3. LC/MS=839.3 (M⁺+1).

Example 173 Preparation of Compound 173

A solution of the thiophen-2-yl-methanol (3.0 mL, 31.7 mmol) in mL ofether was stirred at 0° C. as PBr₃ was added over 5 minutes. After 1.5 hat 0° C., 50% aq. KOH (15 mL) was added and the organic layer wasseparated, which was dried over KOH pellet for 1 h at freezer. The crudesolution of 2-bromomethyl-thiophene was used for the next reaction.

A solution of IV (2.78 g, 8.98 mmol) and DIEA (3.75 mL, 21.53 mmol) in30 mL of CH₂Cl₂ was stirred at r.t. as TMSCl (2.55 mL, 20.09 mmol) wasadded. After 30 min, the crude bromide (16 mL) was added and thesolution was stirred at 40° C. for 16 h. The solution was concentratedand the residue in water (50 mL) was extracted with ethyl acetate (50mL×2). After the organic fraction was washed with water (50 mL), it wasdried (MgSO₄) and concentrated. The residue was purified with acombi-flash column chromatography using hexane:ethyl acetate as eluentto obtain phosphinate (2.191 g, 60%) as a mixture of two diastereomers.

A solution of phosphinate obtained above (2.032 g, 5.01 mmol) in 16 mLof TFA and 4 mL of Me₂S was stirred at r.t. for 6 h. The solution wasconcentrated at 25° C. and the residue in ethyl acetate (60 mL) waswashed with ice-cold 1 N NaOH (60 mL×2) and brine (60 mL). The aqueousfractions were extracted with ethyl acetate (60 mL). The combinedorganic fractions were dried (MgSO₄) and concentrated to obtain 1.22 g(90%) of amine.

To a mixture of VIII (845 mg, 2.31 mmol) and amine (404.8 mg, 1.49 mmol)in toluene (2.7 mL) was added sodium 2-ethyl-hexanoate (164 mg, 0.99mmol) and H₂O (5.5 mL). The reaction mixture was heated to 80° C. andstirred for 19 h. An additional amount of VIII (150 mg) in toluene (1mL) was added and stirred at 80° C. for 7 h. The reaction mixture wascooled to rt, quenched with 1 N HCl (50 mL), and extracted with EtOAc(2×40 mL). The combined organic layers were washed with 1 N HCl (40 mL).The aqueous layer was neutralized with 2 N NaOH (60 mL) beforeextraction with EtOAc (50 mL). The combined organic layers were washedwith saturated NaHCO₃ (2×50 mL) and brine (20 mL), dried with Na₂SO₄,and concentrated. The crude product was purified by combi-flash to give740.5 mg of tripeptide alcohol in 78% yield.

Alcohol (740.8 mg, 1.16 mmol) and DABCO (209.2 mg, 1.87 mmol) weredissolved in toluene (1.1 mL). A solution of brosylchloride (476.8 mg,1.87 mmol) in toluene (0.8 mL) was added dropwise. The reaction mixturewas stirred at r. t. for 2 h. The reaction was diluted with EtOAc (35mL) and washed with 1 N Na₂CO₃. The organic layers were washed with 1 NHCl (30 mL) and H₂O (2×30 mL), dried with Na₂SO₄, filtered, andconcentrated. The crude product was purified by combi-flash to give915.9 mg of brosylate in 92% yield.

Brosylate (915.9 mg, 1.07 mmol) was dissolved in CH₂Cl₂ (150 mL) anddegassed with N₂ for 20 minutes. Grubb's G1 (221.2 mg, 0.27 mmol) wasadded and degassed for an additional 20 minutes. The reaction mixturewas heated to 50° C. overnight and cooled to rt.Tris(hydroxymethyl)phosphine (1.67 g, 3.45 mmol) was added followed byaddition of TEA (3.75 mL, 26.9 mmol) and H₂O (30 mL). The reactionmixture was stirred at rt for 5 h. The two layers were separated. Theorganic layer was washed with H₂O, 0.5 N HCl and saturated NaHCO₃, driedwith Na₂SO₄, and concentrated. The crude product was purified bycombi-flash to give 753.4 mg of cyclic phosphinate in 85% yield.

A solution of cyclic phosphinate (753.4 mg, 0.91 mmol) and2-(2-isopropylamino-thiazol-4-yl)-7-methoxy-quinolin-4-ol (301.3 mg,0.96 mmol) in NMP (10 mL) was treated with Cs₂CO₃ (364.1 mg, 1.12 mmol).The reaction mixture was heated to 70° C. for 12 h and then cooled tort. The reaction mixture was diluted with EtOAc (50 mL) and H₂O (70 mL)and filtered through celite. The two layers were separated. The organiclayer was washed with H₂O (70 mL) and saturated NaHCO₃ (2×70 mL). Theaqueous layer was extracted with EtOAc (50 mL). The combined organiclayers were dried with Na₂SO₄ and concentrated. The crude product waspurified by combi-flash on C18 column to give 470.3 mg of compound.

To a solution of the product obtained above (470.3 mg, 0.52 mmol) inCH₂Cl₂ (3.25 mL) was added 4 N HCl in 1,4-dioxane (3.25 mL, 13 mmol).The reaction mixture was stirred at r.t. for 2.5 h and concentrated. Theresidue was triturated with CH₃CN and concentrated. The residue wasdried under vacuum for 20 minutes to give crude amine HCl salt. Theresulting crude amine HCl salt was dissolved in CH₃CN (5 mL) and cooledto 0° C. Na₂CO₃ (360 mg, 3.4 mmol) in H₂O (5 mL) was added and stirredfor 5 minutes. A solution of the freshly preparedcyclopentylchloroformate in CH₃CN (5 mL, 2.64 mmol) was added. Thereaction was completed within 1 h and concentrated. The residue wasdissolved in EtOAc (40 mL) and washed with H₂O (40 mL) and brine (20mL). The aqueous layers were extracted with EtOAc (20 mL). The combinedorganic layers were dried with Na₂SO₄ and concentrated. The crudeproduct was purified by HPLC to give 286.9 mg of product cyclopetantylcarbamate in 60% yield.

A solution of cyclopetantyl carbamate (87.9 mg, 0.096 mmol) and2,6-lutidine (0.07 mL, 0.6 mmol) in CH₃CN (3 mL) was stirred at 0° C. asiodotrimethylsilane (0.07 mL, 0.49 mmol) was added. The reaction mixturewas warmed to r.t. and stirred for 3.5 h. The reaction mixture wascooled to 0° C. and additional iodotrimethylsilane (0.01 mL) and2,6-lutidine (0.01 mL) were added. The mixture was stirred at r.t. for 1h. MeOH (1 mL) was added and stirred for 1 h. The mixture wasconcentrated and the crude product was purified by HPLC to give 71.2 mgof 173 in 74% yield. ¹H NMR (300 MHz, CD₃OD): d 8.29 (d, 1H, J=9.3 Hz),8.19 (s, 1H), 7.70 (s, 2H), 7.20-7.28 (m, 2H), 6.91-6.97 (m, 2H), 5.79(br, 1H), 5.65 (appt q, 1H, J=˜9 Hz), 5.23 (t, 1H, J=˜9 Hz), 4.88 (d,1H, J=12 Hz), 4.71 (t, 1H, J=8.4 Hz), 4.42 (br, 1H), 4.11-4.24 (m, 2H),4.06 (d, 1H, J=9.9 Hz), 3.99 (s, 3H), 3.64 (t, 1H, J=15.6 Hz), 3.38 (t,1H, J=15.6 Hz), 2.72-2.90 (m, 2H), 2.55-2.66 (m, 1H), 2.17-2.30 (m, 1H),1.81 (br, 2H), 1.17-1.70 (m, 17H), 1.34 (d, 6H, J=6.3 Hz). ³¹P (121.4MHz, CD₃OD): d 38.644. LC/MS=891 (M⁺+1)

Example 174 Preparation of Compound 174

A solution of olefin from Example 173 (193.6 mg, 0.21 mmol) and p-tosylhydrazide (1.26 g, 4.22 mmol) in THF (8 mL) was heated to 60° C. as TEA(0.59 mL, 4.23 mmol) in THF (1 mL) was added in 10 minutes. The reactionmixture was stirred at 60° C. for 27 h. An additional hydrazide (1.26 g,4.22 mmol) and TEA (0.59 mL, 4.23 mmol) were added and stirred for 24 hat 60° C. The reaction was cooled to r.t and filtered off some solid.The filtrate was concentrated and the residue was purified bycombi-flash to give 100.5 mg of a mixture containing the startingmaterial with the desired product. The resulting mixture was dissolvedin THF (5 mL). Hydrazide (655.2 mg, 2.20 mmol) and TEA (0.³¹ mL, 2.22mmol) were added. The mixture was stirred at 60° C. for 20 h. Thereaction mixture was concentrated and the residue was dissolved inCH₂Cl₂. The solid was filtered off and the filtrate was concentrated.The residue was purified by combi-flash to give 68.1 mg of saturatedcompound which was hydrolyzed with 2,6-lutidine (0.1 mL, 0.86 mmol) inCH₃CN (3 mL) and CH₂Cl₂ (1.5 mL) was stirred at 0° C. asiodotrimethylsilane (0.1 mL, 0.7 mmol) was added. The reaction mixturewas warmed to r.t. and stirred for 1.5 h. MeOH (1 mL) was added andstirred at r.t for 0.5 h. The mixture was concentrated and the crudeproduct was purified by HPLC to give 51.3 mg of acid 174. ¹H NMR (300MHz, CD₃OD): d 8.22 (d, 1H, J=9.3 Hz), 8.19 (s, 1H), 7.69 (s, 2H), 7.25(appt d, 2H, J=3.8 Hz), 7.01 (br, 1H), 6.91-6.98 (m, 1H), 5.78 (br, 1H),4.80 (d, 1H, J=11.7 Hz), 4.67 (t, 1H, J=8.4 Hz), 4.45 (br, 1H),4.03-4.27 (m, 3H), 3.98 (s, 3H), 3.57 (t, 1H, J=15.6 Hz), 3.44 (t, 1H,J=15.6 Hz), 2.70-2.82 (m, 1H), 2.42-2.54 (m, 1H), 1.08-1.96 (m, 25H),1.34 (d, 6H, J=6.3 Hz). ³¹P NMR (121.4 MHz, CD₃OD): d 41.072.

LC/MS=893 (M⁺+1)

Example 175 Preparation of Compound 175

A solution of the lactone (VIII) (860 mg, 2.35 mmol), the amine(prepared in Example 83) (300 mg, 1.18 mmol) and sodium 2-ethylhexanoate(60 mg, 0.35 mmol) was stirred in toluene (10 mL) and H₂O (10 mL) at 80°C. for 12 h and then rt for 72 h. Ethyl acetate (50 mL) was added to thesolution and it was washed with saturated sodium carbonate (20 mL), 1MHCl (20 mL) and then brine (15 mL). The organic layer was dried andconcentrated.

To a solution of the resultant diene alcohol (700 mg, 1.13 mmol) andDABCO (200 mg, 1.58 mmol) in toluene (2 mL) was added4-bromotoluenesulfonyl chloride (400 mg, 1.81 mmol) in toluene (5 mL).The solution was stirred at rt for 1.5 h after which the reaction wasdiluted with toluene and quenched with 1M Na₂CO₃. The organic layer waswashed with 1M Na₂CO₃, diluted with ethyl acetate, washed again with0.5M HCl and then H₂O, dried and concentrated. The crude material waspurified by flash chromatography to give the desired brosylate. Thisdiene (290 mg, 0.35 mmol) was placed in degassed CH₂Cl₂ (40 mL) andGrubb's G1 catalyst (7.1 mg, 0.09 mmol) was added. The reaction mixturewas stirred at 40° C. for 15 h. An additional 5 mol % catalyst was addedand the solution continued to stir at 40° C. for 1.5 h. AdditionalGrubb's G1 (5 mol %) was added and the solution continued to stir at 40°C. for 1.5 h. Tris(hydroxymethyl)phosphine (750 g, 1.12 mmol),triethylamine (1.7 mL) and H₂O (25 mL) were added. The solution wasstirred 3 h at 40° C. and then at room temp overnight. The solution wasthen washed with H₂O; the organic layer was dried and concentrated. Thecrude material was purified by HPLC to yield the desired metathesisproduct (264 mg). A solution of this macrocyclic brosylate (260 mg, 0.32mmol), Cs₂CO₃ (104 mg, 0.32 mmol) and aminothiazolequinoline (101 mg,0.32 mmol) in N-methylpyrrolidone (3 mL) was heated at 60° C. for 5 h.The reaction mixture was diluted with ethyl acetate and quenched withsaturated sodium bicarbonate. The organic phase was washed withsaturated bicarbonate solution and brine, then dried and concentrated.The material was purified by column chromatography to provide theproduct (128 mg, 42% over 2 steps).

A solution of the Boc-amine (128 mg, 0.14 mmol) was stirred in CH₂Cl₂ (1mL) as trifluoroacetic acid (500 μl) was added. The solution was stirredat rt for 1 h. The solution was concentrated and azeotroped with toluene(2×). The residue was stirred in THF (1 mL), andcyclopentylchloroformate (0.72 mmol), triethylamine (240 μl, 1.73 mmol)in THF (5001) were added and left at room temp for 1 hour. The reactionmixture was partitioned with H₂O and ethyl acetate, washed with brine,dried (MgSO₄) and concentrated. The crude material was purified with bycolumn chromatography to give the cyclopentyl carbamate (68 mg, 66%).

To a solution of this ethyl phosphinate (68 mg, 0.08 mmol) in CH₃CN (1mL) was added TMSI (54 μl, 0.38 mmol). After 30 minutes, 2,6-lutidine(500 μl) was added, followed by methanol. The reaction mixture wasconcentrated and azeotroped. The crude material was purified by HPLC togive Compound 175 (15 mg, 23%). ¹H NMR (300 MHz, CD₃OD): d 8.31 (d,J=9.4 Hz, 1H), 8.17 (s, 1H), 7.75 (m, 2H), 7.43 (m, 1H), 7.36 (m, 1H),6.36 (m, 1H), 6.28 (m, 1H), 5.85 (m, 1H), 5.73 (m, 1H), 5.28 (m, 1H),4.73 (m, 1H), 4.35 (m, 1H), 4.18 (m, 2H), 4.04 (s, 3H), 2.79 (m, 2H),2.60 (m, 1H), 2.26 (m, 1H) 1.83 (m, 4H), 1.59 (m, 9H), 1.35 (m, 6H). ³¹PNMR (121.4 MHz, CD₃OD):

40.291;

LCMS: 875.39 (M+1).

Example 176 Preparation of Compound 176

The methathesis product from Example 175 (1.2 g, 1.48 mmol) was taken upin ethyl acetate (15 mL) and Degussa-type Rhodium on Alumina (0.6 g, 50%w/w) was added. The reaction vessel was evacuated and placed underhydrogen atmosphere, stirred at rt for 5 h. When the reaction wascomplete, the solution was filtered through celite and concentrated toprovide 1.08 g which was used directly in the next step.

This brosylate (1.08 g, 1.33 mmol), was taken up inN-methylpyrrolidinone (15 mL) and the aminothiazolequinoline (0.42 g,1.33 mmol) and cesium carbonate (0.43 g, 1.33 mmol) were added. Thereaction mixture was heated at 60° C. for 4 h, and then stirred at roomtemperature overnight. Ethyl acetate was added and the organic layer waswashed with saturated sodium bicarbonate solution, dried, andconcentrated. The residue was purified by flash chromatography toprovide the desired product (0.45 g, 38% in two steps).

This Boc-amine (0.45 g, 0.5 mmol) was taken up in acetonitrile (5 mL),and TMSI (0.36 mL, 2.5 mmol) was added. The reaction mixture was stirredat room temperature for 15 minutes (LCMS analysis showed fullconversion). 2,6-Lutidine (0.2 mL) was added, followed by quenching withmethanol, concentrating and azeotroping with toluene (3×20 mL). Theresidue was then taken up in acetonitrile/water (5 mL each). Sodiumcarbonate (0.64 g, 6 mmol) and cyclopentyl chloroformate (5 equiv.) wereadded, and the reaction mixture was stirred at room temperature for 1.5h. LCMS showed full conversion and the reaction was then concentrated,filtered, and purified by HPLC to provide compound 176 (180 mg, 41%). ¹HNMR (300 MHz, CD₃OD)

8.30 (d, 1H J=9.4 Hz), 8.07 (s, 1H), 7.75 (s, 2H), 7.43 (m, 1H), 7.35(m, 1H), 6.37 (m, 1H), 6.29 (m, 1H), 5.83 (m, 1H), 4.79 (min, 1H), 4.65(m, 1H), 4.41 (m, 1H), 4.18 (m, 3H), 4.04 (s, 3H), 2.79 (m, 1H), 2.51(m, 1H), 1.89 (m, 2H), 1.76 (m, 2H), 1.42 (m, 24H), 1.34 (d, 6H, J=6.7Hz). ³¹P NMR (121.4 MHz, CD₃OD):

2.676. LCMS: 877.16 (M+1).

Example 177 Preparation of Compound 177

A solution of cyclic brosylate (Example 173) (448.5 mg, 0.54 mmol) and7-methoxy-2-phenyl-quinolin-4-ol (145 mg, 0.58 mmol) in NMP (6.8 mL) wastreated with Cs₂CO₃ (224 mg, 0.69 mmol). The reaction mixture was heatedto 70° C. for 8 h and then cooled to rt. The reaction mixture wasdiluted with EtOAc (80 mL) and washed with H₂O (2×100 mL) followed bysaturated NaHCO₃ (2×100 mL). The aqueous layer was extracted with EtOAc(80 mL). The combined organic layers were dried with Na₂SO₄ andconcentrated. The crude product was purified by combi-flash to give295.7 mg of desired product in 65% yield.

To a solution of product obtained above (295.7 mg, 0.35 mmol) in CH₂Cl₂(2.2 mL) was added 4 N HCl in 1,4-dioxane (2.2 mL, 8.8 mmol). Thereaction mixture was stirred at r.t. for 2 h and concentrated. Theresidue was triturated with CH₃CN and concentrated. The residue wasdried under vacuum for 20 minutes to give crude amine HCl salt.Cyclopentanol (0.16 mL, 1.76 mmol) was dissolved in THF (6 mL) and 20%phosgene in toluene (1.5 mL, 2.84 mmol) was added. The reaction wasstirred for 1 h and concentrated. The residue was dissolved in CH₂Cl₂and concentrated to give crude cyclopentylchloroformate. The crude amineHCl salt was dissolved in CH₃CN (2 mL) and cooled to 0° C. Na₂CO₃ (225mg, 2.12 mmol) in H₂O (3 mL) was added and stirred for 5 minutes. Asolution of the freshly prepared cyclopentylchloroformate in CH₃CN (4mL) was added. The reaction was completed within 1 h and concentrated.The residue was dissolved in EtOAc (30 mL) and washed with H₂O (30 mL)and brine (20 mL). The aqueous layer was extracted with EtOAc (20 mL).The combined organic layers were dried with Na₂SO₄ and concentrated. Thecrude product was purified by combi-flash to give 255.9 mg of product.

A solution of product obtained above (97.5 mg, 0.11 mmol) and2,6-lutidine (0.08 mL, 0.69 mmol) in CH₃CN (2 mL) was stirred at 0° C.as iodotrimethylsilane (0.08 mL, 0.56 mmol) was added. The reactionmixture was stirred at 0° C. for 6 h and then warmed to r.t for 30minutes. MeOH (1 mL) was added and stirred for 20 minutes. The mixturewas concentrated and the crude product was purified by HPLC to give 96.1mg of acid 177 in 90% yield. ¹H NMR (300 MHz, CD₃OD): d 8.43 (d, 1H,J=9.3 Hz), 8.09 (dd, 2H, J=8.0 and 1.7 Hz), 7.71-7.82 (m, 3H), 7.67 (s,1H), 7.55 (d, 1H, J=2.4 Hz), 7.38 (dd, 1H, J=9.3 and 2.4 Hz), 7.18-7.23(m, 1H), 6.90-6.96 (m, 2H), 5.89 (br, 1H), 5.64 (appt q, 1H, J=˜9 Hz),5.24 (appt t, 1H, J=9.9 Hz), 4.96 (1H), 4.74 (dd, 1H, J=9.3 and 7.5 Hz),4.42 (br, 1H), 4.18 (d, 1H), 4.07 (d, 1H), 4.03 (s, 3H), 3.62 (t, 1H,J=15.3 Hz), 3.36 (t, 1H, J=15.3 Hz), 2.74-2.88 (m, 2H), 2.55-2.67 (m,1H), 2.17-2.29 (m, 1H), 1.80 (brm, 1H), 1.16-1.69 (m, 18H). ³¹P NMR(121.4 MHz, CD₃OD): d 38.060. LC/MS=827 (M⁺+1)

Example 178 Preparation of Compound 178

A solution of ethyl phosphinate (Example 177, 158.1 mg, 0.18 mmol) andp-tosyl hydrazide (551.9 mg, 1.85 mmol) in THF (6 mL) was heated to 70°C. as TEA (0.26 mL) in THF (1 mL) was added in 10 minutes. The reactionmixture was stirred at 60° C. overnight. An additional hydrazide (555mg) and TEA (0.26 mL) in THF (1 mL) were added, and stirred for 16 h at60° C. More hydrazide (552 mg) and TEA (0.26 mL) in THF (1 mL) wereadded, and stirred at 60° C. for 9 h. The mixture was stored in freezerfor 5.5 days. Hydrazide (1.11 g) and TEA (0.52 mL) were added, andstirred at 60° C. for 20 h. The reaction mixture was concentrated andthe residue was diluted with EtOAc before washing with 0.5 N NaOH (2×50mL). The aqueous layer was extracted with EtOAc (2×25 mL). The organiclayers were washed with brine, dried with Na₂SO₄, and concentrated. Theresidue was purified by combi-flash to give 88.3 mg of a mixturecontaining 10% of product. The resulting mixture was dissolved in CH₃CN(5 mL). Hydrazide (618 mg, 2.07 mmol) and TEA (0.29 mL) were added. Themixture was stirred at 60° C. overnight. The reaction mixture wasconcentrated and the residue was dissolved in EtOAc (30 mL). Thesolution was washed with ice cold 1 N NaOH (2×20 mL) and brine. Theaqueous layer was extracted with EtOAc (2×20 mL). The combined organiclayers were dried with Na₂SO₄ and concentrated. The residue was purifiedby thin layer chromatography to give 71.5 mg of saturated compound.

A solution of saturated compound (71.5 mg, 0.08 mmol) and 2,6-lutidine(0.06 mL, 0.52 mmol) in CH₃CN (2 mL) was stirred at 0° C. asiodotrimethylsilane (0.06 mL, 0.42 mmol) was added. The reaction mixturewas warmed to r.t. and stirred for 2 h. MeOH (1 mL) was added andstirred for 1 h. The mixture was concentrated and the crude product waspurified by HPLC to give 49.2 mg of acid 178. ¹H NMR (300 MHz, CD₃OD): d8.36 (d, 1H, J=9.3 Hz), 8.09 (dd, 2H, J=8.1 and 1.5 Hz), 7.68-7.81 (m,3H), 7.66 (s, 1H), 7.56 (d, 1H, J=2.1 Hz), 7.37 (dd, 1H, J=9.3 and 2.1Hz), 7.24 (d, 1H, J=4.8 Hz), 7.00 (br, 1H), 6.95 (dd, 1H, J=4.8 and 3.3Hz), 5.89 (br, 1H), 4.87 (d, 1H, J=˜12 Hz), 4.71 (t, 1H, J=8.4 Hz), 4.43(br, 1H), 4.24 (m, 1H), 4.09 (m, 1H), 4.02 (s, 3H), 3.56 (t, 1H, J=15.6Hz), 3.43 (t, 1H, J=15.6 Hz), 2.81 (dd, 1H, J=14.1 and 7.5 Hz),2.44-2.56 (m, 1H), 1.10-1.96 (m, 25H). ³¹P NMR (121.4 MHz, CD₃OD): d41.123. LC/MS=829 (M⁺+1)

Example 179 Preparation of Compound 179

A solution of the brosylate from Example 175 (255 mg, 0.31 mmol), Cs₂CO₃(102 mg, 0.3 mmol) and phenylquinoline (79 mg, 0.31 mmol) stirred inN-methylpyrrolidone (3 mL) at 60° C. After 4 h, the reaction was dilutedwith ethyl acetate and quenched with saturated NaHCO₃. The organic layerwas washed with saturated NaHCO₃ (2×) and brine. The organic layer wasdried and concentrated. The material was purified by columnchromatography to provide the desired product (114 mg, 41%).

A solution of the Boc-amine (114 mg, 0.14 mmol) and trifluoroacetic acid(500 μl) in CH₂Cl₂ (1 mL) was stirred at rt for 1 h. The solution wasthen concentrated, and azeotroped with toluene (2×). The residue wasstirred in THF (1 mL), and cyclopentyl chloroformate (0.69 mmol) andtriethylamine (230 μl, 1.66 mmol) in THF (500 μl) were sequentiallyadded. The solution was stirred at rt for 1 hour. The reaction mixturewas partitioned between water and ethyl acetate. The organic layer waswashed with brine, dried (MgSO₄) and concentrated. The crude materialwas purified by column chromatography to give the ethyl phosphinate (86mg, 76%).

A solution of the ethyl phosphinate (39 mg, 0.04 mmol) and TMSI (30 μl,0.23 mmol) in CH₃CN (1 mL) was stirred for 30 minutes, and then2,6-Lutidine (500 μl) was added. The solution was concentrated andazeotroped with methanol and toluene. The crude material was purified byHPLC to give compound 179 (21 mg, 56%). ¹H NMR (300 MHz, CD₃OD): d 8.40(d, 1H, J=9.4 Hz), 8.08 (m, 1H), 7.73 (m, 5H), 7.52 (m, 1H), 7.38 (m,2H), 6.33 (m, 1H), 6.22 (m, 1H), 5.91 (m, 1H), 5.73 (m 1H), 5.26 (m,1H), 4.71 (m, 1H), 4.35 (m, 1H), 4.15 (m, 1H), 5.24 (s, m, 4H), 2.80 (m,2H), 2.62 (m, 1H), 2.26 (m, 1H), 1.81 (m, 2H), 1.50 (m, 14H). ³¹P NMR(121.4 MHz) d 40.245. LCMS. 812 (M+1).

Example 180 Preparation of Compound 180

Following experimental procedures similar to those described for thepreparation of compound 152, compound 180 was prepared. ¹H NMR (300 MHz,CD₃OD) d 8.34 (d, J=9.6 Hz, 1H), 8.31 (s, 1H), 7.86 (s, 1H), 7.65 (d,J=9.6 Hz, 1H), 5.84 (bs, 1H), 4.84 (m, 1H), 4.67 (t, J=7.5 Hz, 1H), 4.33(bs, 1H), 4.17 (m, 1H), 4.16 (s, 3H), 4.03 (m, 2H), 3.47 (m, 2H), 2.83(m, 1H), 2.58 (m, 1H), 2.0-2.3 (m, 2H), 1.6-2.0 (m, 2H), 1.2-1.6 (m,3H); ³¹P NMR (121.4 MHz, CD₃OD)

38.831; LCMS (M+1): 956.2.

Example 181 Preparation of Compound 181

A solution of brosylate (Example 171) (182 mg, 0.23 mmol), Cs₂CO₃ (76mg, 0.23 mmol) and 7-Methoxy-2-phenyl-quinolin-4-ol (58 mg, 0.23 mmol)in 1-methyl-2-pyrrolidinone (2.3 mL) was stirred at 65° C. for 3.5 h.The solution was diluted with EtOAc and quenched with 2.5% NaHCO₃solution. The organic phase was washed with 2.5% NaHCO₃ solution (2×)and brine, dried over MgSO₄, and concentrated.

A solution of crude product (93 mg, 0.12 mmol) and 2,6-lutidine (81 μl,0.70 mmol) in CH₃CN (3 mL) was stirred at 0° C. as iodotrimethylsilane(99 μl, 0.70 mmol) was added drop-wise. The solution was warmed to rtand stirred for 1.5 h. The solution was cooled back to 0° C., andadditional 2,6-Lutidine (40 μl, 0.35 mmol) and iodotrimethylsilane (49μl, 0.35 mmol) were added. The solution was then stirred again at rt for1.5 h. The solution was cooled to 0° C., and Et₃N (500 μl) and MeOH (1mL) were added. The reaction mixture was concentrated and 181 (70 mg,78%) was obtained from the residue by HPLC purification as a whitesolid. ¹H NMR (300 MHz, CD₃OD):

8.42 (d, J=9.3 Hz, 1H), 8.08 (d, J=6.9 Hz, 2H), 7.76 (m, 3H), 7.66 (s,1H), 7.55 (s, 1H), 7.35 (d, J=9.3 Hz, 1H), 5.88 (s, 1H), 5.54 (q, J=9Hz, 1H), 5.18 (t, J=9.6 Hz, 1H), 4.9 (s, 1H), 4.73 (t, J=8.7 Hz, 1H),4.42 (s, 1H), 4.17 (d, J=11.4 Hz, 1H), 4.07 (d, J=11.7 Hz, 1H), 4.02 (s,3H), 2.80 (m, 2H), 2.58 (m, 1H), 2.20 (m, 1H), 1.2-2.0 (brm, 2H), 1.00(t, J=7.2 Hz, 3H). ³¹P NMR (121.4 MHz, CD₃OD): d 46.5

LC/MS=773.8 (M⁺+1).

Example 182 Preparation of Compound 182

A solution of phosphinate (Example 181, 81 mg, 0.10 mmol) and2,4,6-triisopropylbenzene-sulfonylhydrazide (292 mg, 0.98 mmol) in THF(3 mL) was stirred at 60° C. as Et₃N (137 g, 0.98 mmol) was addeddropwise over 5 minutes. After 1 h, the reaction was cooled andadditional 2,4,6-triisopropylbenzene-sulfonylhydrazide (292 mg, 0.98mmol) was added. This was followed by the slow addition of more Et₃N(137 μl, 0.98 mmol). After 1 h, more2,4,6-triisopropylbenzene-sulfonylhydrazide (292 mg, 0.98 mmol) and Et₃N(137 μl, 0.98 mmol) was added. After 1 h, the reaction was diluted withEtOAc and washed with saturated NH₄Cl (2×). THF was added to the organiclayer and it was extracted with saturated NH₄Cl, saturated Na₂CO₃, andH₂O. The resulting mono-phasic solution was extracted with EtOAc.Combined organic washes were extracted with brine, and dried over MgSO₄and concentrated. Saturated phosphinate (70 mg, 87%) was purified fromthe residue by HPLC to yield a glassy white solid. LC/MS=802.7 (M⁺+1),824.4 (M⁺+Na)

A solution of saturated phosphinate (70 mg, 0.09 mmol) and 2,6-lutidine(61 μl, 0.52 mmol) in CH₃CN (3 mL) was cooled to 0° C. andiodotrimethylsilane (75 μl, 0.52 mmol) was added drop-wise. The solutionwas stirred at rt for 1 h. The reaction was then cooled to 0° C. andadditional 2,6-lutidine (30 μl, 0.26 mmol) and iodotrimethylsilane (37μl, 0.26 mmol) was added. The solution then warmed again to rt andstirred for 3 h. The reaction was cooled to 0° C. and Et₃N (500 μl) andMeOH (1 mL) was added. The reaction was concentrated and 182 (46 mg,69%) was isolated from the residue by HPLC as an off-white solid.

¹H NMR (300 MHz, CD₃OD)

8.37 (d, J=9 Hz, 1H), 8.08 (m, 2H), 7.76 (m, 3H), 7.66 (s, 1H), 7.55 (s,1H), 7.39 (dd, J=1.8, 9.3 Hz, 1H), 5.89 (1H), 4.69 (t, J=9 Hz, 1H), 4.43(s, 1H), 4.24 (d, J=7.5 Hz, 1H), 4.09 (d, J=12 Hz, 1H), 4.04 (s, 3H),2.82 (dd, J=7.2, 14.4 Hz), 2.50 (ddd, J=4.2, 9.3, 13.8 Hz, 1H), 1.2-2.0(brm, 28H), 1.05 (t, J=7.2 Hz, 3H). ³¹P NMR (121.4 MHz, CD₃OD): d 50.36.LC/MS=775.3 (M⁺+1)

Example 183 Preparation of Compound 183

Step 1. To a solution of the BOC-protected phosphinate (described inExample 143, 2.22 g, 2.46 mmol) in CH₂Cl₂ (10 mL) was added 4 N HCl in1,4-dioxane (50 mL, 200 mmol). The reaction mixture was stirred at r.t.for 3.5 h, concentrated, co-evaporated with CH₂Cl₂, and dried undervacuum to give the desired amine as a brown solid.

Step 2. To a solution of the amine obtained from step 1 (60 mg, 0.075mmol) in CH₂Cl₂ (0.7 mL) at 0° C. was added 6.8 M cyclopentylisocyanatein CH₂Cl₂ (0.2 mL, 0.062 mmol). The reaction mixture was stirred for 1h, kept in −20° C. freezer overnight, and concentrated. The residue waspurified by combi-flash to give 50.6 mg of intermediate ester. Theresulting ester (50 mg) was dissolved in CH₃CN (2 mL) and cooled to 0°C. Iodotrimethylsilane (0.2 mL) was added. The reaction mixture waswarmed to rt, stirred for 0.5 h, and cooled to 0° C. TEA (0.5 mL) wasadded followed by addition of MeOH (2 mL). The mixture was concentratedand purified by HPLC to give 11.6 mg of compound 183. ¹H NMR (300 MHz,CD₃OD):

8.35 (d, J=9.3 Hz, 1H), 8.19 (s, 1H), 7.74 (s, 2H), 7.15-7.42 (m, 6H),5.82 (brs, 1H), 5.68 (dd, J=8.6, 18.3 Hz, 1H), 5.27 (t, J=9.6 Hz, 1H),4.83 (d, J=12.0 Hz, 1H), 4.72 (t. J=8.7 Hz, 1H), 4.³¹ (d, J=8.7 Hz, 1H),4.07-4.23 (m, 2H), 4.03 (s, 3H), 3.51 (m, 1H), 3.38 (t, J=15.9 Hz, 1H),3.17 (t, J=15.9 Hz, 1H), 2.79 (m, 1H), 2.61 (m, 1H), 2.25 (m, 1H),1.2-1.9 (m, 26H); ³¹P NMR (121.4 MHz, CD₃OD):

42.114; LC/MS=884 (M⁺+1).

Example 184 Preparation of Compound 184

4-Trifluorobutyric acid (15.5 mg, 0.11 mmol) and the amine HCl salt(Example 183, 70 mg, 0.084 mmol) were dissolved in CH₂Cl₂ (1 mL)/DMF(0.5 mL). HATU (47.88 mg, 0.13 mmol) and DIPEA (0.06 mL, 0.34 mmol) wereadded and the mixture was stirred at r.t. for 15 minutes. The reactionmixture was concentrated, re-dissolved in EtOAc, washed with 5% LiCl,saturated NaHCO₃ and brine, and concentrated. The crude product waspurified by combi-flash to give 33.4 mg of intermediate ester. Theresulting ester (33.4 mg, 0.036 mmol) was dissolved in CH₃CN (2 mL) andcooled to 0° C. Iodotrimethylsilane (0.2 mL) was added. The reactionmixture was warmed to rt, stirred for 0.5 h, and cooled to 0° C. TEA(0.5 mL) was added followed by addition of MeOH (2 mL). The mixture wasconcentrated and purified by HPLC to give 17.7 mg of compound 184. ¹HNMR (300 MHz, CD₃OD):

8.29 (d, J=9.3 Hz, 1H), 8.18 (s, 1H), 7.75 (s, 2H), 7.15-7.42 (m, 6H),5.85 (brs, 1H), 5.73 (dd, J=8.6, 18.3 Hz, 1H), 5.30 (t, J=9.6 Hz, 1H),4.69 (dd, J=7.5, 9.3 Hz, 1H), 4.30 (d, J=9.6 Hz, 1H), 4.05-4.22 (m, 2H),4.04 (s, 3H), 3.51 (m, 1H), 3.39 (t, J=15.3 Hz, 1H), 3.18 (t, J=15.3 Hz,1H), 2.81 (m, 2H), 2.61 (m, 1H), 2.25 (m, 3H), 2.02 (m, 2H), 1.50 (m,3H), 1.4-1.6 (m, 8H), 1.34 (d, J=6.3 Hz, 6H); ³¹P NMR (121.4 MHz,CD₃OD):

2.563; LC/MS=897 (M⁺+1).

Example 185 Preparation of Compound 185

Following experimental procedures similar to those described for thepreparation of compound 184 compound 185 was prepared. ¹H NMR (300 MHz,CD₃OD):

8.29 (d, J=9.3 Hz, 1H), 8.19 (s, 1H), 7.76 (s, 2H), 7.15-7.38 (m, 6H),5.84 (brs, 1H), 5.73 (dd, J=8.6, 18.3 Hz, 1H), 5.29 (t, J=9.6 Hz, 1H),4.94 (d, J=12.0 Hz, 1H), 4.69 (t, J=8.7 Hz, 1H), 4.36 (d, J=8.7 Hz, 1H),4.07-4.23 (m, 2H), 4.05 (s, 3H), 3.38 (t, J=15.9 Hz, 1H), 3.18 (t,J=15.9 Hz, 1H), 2.82 (m, 2H), 2.61 (m, 1H), 2.25 (m, 1H), 2.02 (d, J=7.5Hz, 2H), 1.85 (m, 3H), 1.2-1.9 (m, 23H); ³¹P NMR (121.4 MHz, CD₃OD):

42.453; LC/MS=883 (M⁺+1).

Example 186 Preparation of Compound 186

Following experimental procedures similar to those described for thepreparation of compound 184, compound 186 was prepared. LC/MS=909(M⁺+1).

Example 187 Preparation of Compound 187

To a solution of amine (Example 185) (130 mg, 0.16 mmol) andBoc-L-tert-leucine (45 mg, 0.20 mmol) were dissolved in CH₂Cl₂ (2mL)/DMF (0.5 mL). HATU (93 mg, 0.24 mmol) and DIPEA (0.13 mL, 0.45 mmol)were added and the mixture was stirred at r.t. for 15 minutes. Thereaction mixture was concentrated, re-dissolved in EtOAc, washed with 5%LiCl, saturated NaHCO₃ and brine, and concentrated. The crude productwas purified by combi-flash to give 133 mg of tetrapeptide in 81% yield.

The tetrapeptidic intermediate (133 mg, 0.13 mmol) was dissolved inCH₃CN (2 mL) and cooled to 0° C. Iodotrimethylsilane (0.2 mL) was added.The reaction mixture was warmed to rt, stirred for 0.5 h, and cooled to0° C. TEA (0.4 mL) was added followed by addition of MeOH (2 mL). Themixture was concentrated and purified by HPLC to give 82.6 mg of 187 in57% yield. ¹H NMR (300 MHz, CD₃OD):

8.24 (d, J=9.3 Hz, 1H), 8.21 (s, 1H), 7.78 (s, 2H), 7.36 (dd, J=2.4, 9.0Hz, 1H) 7.15-7.38 (m, 5H), 5.86 (brs, 1H), 5.78 (dd, J=8.6, 18.3 Hz,1H), 5.33 (t, J=9.6 Hz, 1H), 4.86 (d, J=12.0 Hz, 1H), 4.67 (t, J=8.7 Hz,1H), 4.54 (d, J=8.7 Hz, 1H), 4.1-4.25 (m, 2H), 4.06 (s, 3H), 3.47 (s,1H), 3.36 (t, J=15.9 Hz, 1H), 3.19 (t, J=15.9 Hz, 1H), 2.82 (m, 2H),2.61 (m, 1H), 2.32 (m, 1H), 2.02 (d, J=7.5 Hz, 1H), 1.98 (m, 2H),1.23-1.7 (m, 8H), 1.34 (d, J=6.3 Hz, 6H), 0.83 (s, 9H); ³¹P NMR (121.4MHz, CD₃OD):

42.485; LC/MS=886 (M⁺+1).

Example 188 Preparation of Compound 188

To a mixture of the amine prepared as shown in Example 185 (60 mg, 0.075mmol) and p-tolylboronic acid (20 mg, 0.15 mmol) in CH₂Cl₂ (4 mL) wasadded molecular sieves (150 mg), TEA (0.2 mL), and Cu(OAc)₂sequentially. The reaction mixture was stirred under air with dryingtube for 18 h, diluted with CH₂Cl₂, and filtered through celite. Thefiltrate was concentrated and purified by HPLC to give 100 mg ofintermediate ester. The ester (100 mg, 0.075 mmol) and 2,6-lutidine(0.09 mL, 0.75 mmol) were dissolved in CH₃CN (2 mL) and cooled to 0° C.Iodotrimethylsilane (0.05 mL, 0.38 mmol) was added. The reaction mixturewas warmed to rt, stirred for 1 h, and cooled to 0° C. MeOH (0.2 mL) wasadded and warmed to rt. The mixture was concentrated and purified byHPLC to give 4.6 mg of 188 as a yellow solid. ¹H NMR (300 MHz, CD₃OD):

8.19 (s, 1H

7.96 (s, 1H), 7.81 (s, 1H), 7.74 (s, 1H), 7.25-7.35 (m, 6H), 6.61 (m,4H), 5.84 (brs, 1H), 5.78 (dd, J=8.6, 18.3 Hz, 1H), 5.36 (t, J=9.6 Hz,1H), 4.64 (t, J=8.7 Hz, 1H), 4.30-4.47 (m, 2H), 4.10-4.25 (m, 2H), 4.08(s, 3H), 3.39 (t, J=15.9 Hz, 1H), 3.18 (t, J=15.9 Hz, 1H), 2.75 (m, 1H),2.63 (m, 1H), 2.28 (m, 1H), 1.98 (s, 3H), 1.7-2.1 (m, 3H), 1.2-1.7 (m,14H); ³¹P NMR (121.4 MHz, CD₃OD):

42.409; LC/MS=863 (M⁺+1).

Example 189 Preparation of Compound 189

Following experimental procedures similar to those described for thepreparation of compound 184, compound 189 was prepared. ¹H NMR (300 MHz,CD₃OD):

20 (s, 1H

8.13 (d, J=9.0 Hz, 1H), 7.82 (s, 1H), 7.80 (s, 1H), 7.20-7.38 (m, 6H),6.88 (d, J=8.1 Hz, 2H), 6.50 (d, J=8.1 Hz, 2H), 5.94 (brs, 1H), 5.77(dd, J=8.6, 18.3 Hz, 1H), 5.36 (t, J=9.6 Hz, 1H), 4.74 (t, J=8.7 Hz,1H), 4.60 (d, J=12.3 Hz, 1H), 4.43 (m, 1H), 4.10-4.25 (m, 2H), 4.07 (s,3H), 3.36 (t, J=15.9 Hz, 1H), 3.20 (t, J=15.9 Hz, 1H), 2.63-2.82 (m,2H), 2.24 (m, 1H), 1.7-2.1 (m, 3H), 1.2-1.7 (m, 14H); ³¹P NMR (121.4MHz, CD₃OD):

42.592; LC/MS=917 (M⁺+1).

Example 190 Preparation of Compound 190

Following experimental procedures similar to those described for thepreparation of compound 184, compound 190 was prepared. ¹H NMR (300 MHz,CD₃OD):

8.18 (s, 1H

7.99 (d, J=9.3 Hz, 1H), 7.76 (s, 1H), 7.75 (s, 1H), 7.15-7.33 (m, 6H),6.90 (m, 2H), 6.78 (d, J=7.5 Hz, 1H), 6.72 (d, J=7.5 Hz, 1H), 5.89 (brs,1H), 5.75 (dd, J=8.6, 18.3 Hz, 1H), 5.34 (t, J=9.6 Hz, 1H), 4.72 (t,J=8.7 Hz, 1H), 4.49 (brs, 1H), 4.44 (d, J=12.3 Hz, 1H), 4.10-4.30 (m,2H), 4.07 (s, 3H), 3.35 (t, J=15.9 Hz, 1H), 3.19 (t, J=15.9 Hz, 1H),2.63-2.82 (min, 2H), 2.24 (m, 1H), 1.7-2.1 (m, 3H), 1.2-1.7 (m, 14H);³¹P NMR (121.4 MHz, CD₃OD):

42.205; LC/MS=917 (M⁺+1).

Example 191 Preparation of Compound 191

Following experimental procedures similar to those described for thepreparation of compound 184, compound 191 was prepared. ¹H NMR (300 MHz,CD₃OD):

8.16 (s, 1H

8.12 (d, J=9.3 Hz, 1H), 7.75 (s, 1H), 7.74 (s, 1H), 7.15-7.33 (m, 6H),7.08 (t, J=7.2 Hz, 1H), 6.81 (d, J=7.2 Hz, 1H), 6.63 (d, J=7.2 Hz, 1H),5.88 (brs, 1H), 5.75 (dd, J=8.6, 18.3 Hz, 1H), 5.40 (t, J=9.6 Hz, 1H),4.78 (t, J=8.7 Hz, 1H), 4.64 (m, 1H), 4.39 (d, J=12.3 Hz, 1H), 4.10-4.30(m, 2H), 4.06 (s, 3H), 3.35 (t, J=15.9 Hz, 1H), 3.21 (t, J=15.9 Hz, 1H),2.63-2.82 (m, 2H), 2.24 (m, 1H), 1.7-2.1 (m, 3H), 1.2-1.7 (m, 14H); ³¹PNMR (121.4 MHz, CD₃OD): d 42.896; LC/MS=917 (M⁺+1).

Example 192 Preparation of Compound 192

To a solution of the amine prepared as step 1 of Example 185 (70 mg,0.087 mmol) in THF (2 mL) was added thiocarbonyl diimidazole and stirredat r.t. for 1 h. 2.0 M ammonium in MeOH (1 mL) was added and stirred at50° C. for 1 h in a capped vial and concentrated. The residue wasdissolved in CH₂Cl₂ (2 mL), the a-bromoketone (35 μL) was added, stirredat 50° C. for 1 h, and concentrated. The crude product was purified bycombi-flash to give 80 mg of the phosphinate. Deprotection withiodotrimethylsilane (0.1 mL) provided 192. ¹H NMR (300 MHz, CD₃OD):

8.20 (S, 1H

8.16 (d, J=9.3 Hz, 1H), 7.80 (s, 1H), 7.77 (s, 1H), 7.73 (s, 1H),7.18-7.38 (m, 6H), 5.95 (brs, 1H), 5.79 (dd, J=8.6, 18.3 Hz, 1H), 5.39(t, J=9.6 Hz, 1H), 4.82 (t, J=8.7 Hz, 1H), 4.72 (brs, 1H), 4.44 (d,J=12.3 Hz, 1H), 4.24 (dd, J=12.6, 3.3 Hz, 1H), 4.17 (m, 1H), 4.03 (s,3H), 3.39 (t, J=15.9 Hz, 1H), 3.18 (t, J=15.9 Hz, 1H), 2.82 (m, 1H),2.61 (m, 1H), 2.24 (m, 1H), 1.91 (m, 2H), 1.65 (m, 1H), 1.2-1.7 (m,23H); ³¹P NMR (121.4 MHz, CD₃OD):

42.306.

Example 193 Preparation of Compound 193

A mixture of compound 184 (308 mg, 0.305 mmol), tosyl hydride (425 mg,2.28 mmol) and sodium acetate (375 mg, 4.58 mmol) in a mixed solvents of1,2-dimethoxyethane (5.5 mL) and water (0.6 mL) was stirred at 95° C.for 3 h. The reaction mixture was then concentrated to a volume of 3 mLand filtered. The filtrate was purified by HPLC, affording 250 mg (81%)of compound 193. ¹H NMR (300 MHz, CD₃OD):

8.27 (d, J=9.6 Hz, 1H), 8.17 (s, 1H), 7.76 (s, 2H), 7.2-7.4 (m, 6H),5.84 (brs, 1H), 4.83 (d, J=12.3 Hz, 1H), 4.65 (t, J=8.7 Hz, 1H), 4.39(m, 1H), 4.24 (brs, 1H), 4.04-4.23 (m, 3H), 4.15 (m, 2H), 3.39 (t,J=15.9 Hz, 1H), 3.28 (t, J=15.9 Hz, 1H), 2.78 (m, 1H), 2.49 (m, 1H),2.30 (m, 2H), 1.7-2.1 (m, 5H), 1.1-1.7 (m, 19H); ³¹P NMR (121.4 MHz,CD₃OD):

44.910; LC/MS=899 (M⁺+1).

Example 194 Preparation of Compound 194

Following experimental procedures similar to those described for thepreparation of compound 184, compound 194 was prepared. ¹H NMR (300 MHz,CD₃OD):

3.26 (d, J=9.3 Hz, 1H), 8.18 (s, 1H), 7.75 (s, 2H), 7.20-7.35 (m, 6H),5.82 (brs, 1H), 4.84 (d, J=12.3 Hz, 1H), 4.64 (t, J=8.7 Hz, 1H), 4.44(m, 1H), 4.15 (m, 3H), 4.05 (s, 3H), 3.34 (t, J=15.9 Hz, 1H), 3.22 (t,J=15.9 Hz, 1H), 2.77 (m, 1H), 2.48 (m, 1H), 2.04 (d, 2H), 1.7-2.0 (m,3H), 1.1-1.7 (m, 26H), 1.00 (m, 2H); ³¹P NMR (121.4 MHz, CD₃OD):

44.979; LC/MS=885 (M⁺+1).

Example 195 Preparation of Compound 195

The fully protected macrocyclic phosphinate (synthesized as described inExample 152 with the Boc protection group) was treated with HCl toremove the Boc protection group. The resulting amine was used to preparecompounds 195-200.

To a solution of this amine (34 mg, 0.04 mmol) in EtOAc (2 mL) was addedsaturated NaHCO₃ (2 mL) and stirred vigorously. 2,2-Dimethylpropylchloroformate (7 μL) was added and stirred for 15 minutes. The twolayers were separated. The organic layer was washed with brine andconcentrated. The dried residue was dissolved in CH₃CN (1 mL) and cooledto 0° C. Iodotrimethylsilane (0.20 mL) was added. The reaction mixturewas stirred for 0.5 h at 0° C. 2,6-lutidine (0.6 mL) was added followedby addition of MeOH (3 mL). The mixture was concentrated in vacuo. Theresidue was purified by HPLC to give 25.2 mg of compound 195.LC/MS=959.1 (M⁺+1).

Example 196 Preparation of Compound 196

Following experimental procedures similar to those described for thepreparation of compound 184, compound 196 was prepared. LC/MS=915.2(M⁺+1).

Example 197 Preparation of Compound 197

Following experimental procedures similar to those described for thepreparation of compound 184, compound 197 was prepared. LC/MS=955.1(M⁺+1).

Example 198 Preparation of Compound 198

Following experimental procedures similar to those described for thepreparation of compound 184, compound 198 was prepared. LC/MS=983.1(M⁺+1).

Example 199 Preparation of Compound 199

Following experimental procedures similar to those described for thepreparation of compound 184, compound 199 was prepared. LC/MS=944.3(M⁺+1).

Example 200 Preparation of Compound 200

Compound 200 was prepared using a method similar to that described forcompound 194. LCMS (M+1): 955.24.

Example 201 Preparation of Compound 201

Compound 201 was prepared using a method similar to that described forcompound 177. LCMS (M+1): 821.36.

Example 202 Preparation of Compound 202

Compound 202 was prepared using a method similar to that described forcompound 178. LCMS (M+1): 823.37.

Example 203 Preparation of Compound 203

Compound 203 was prepared using the same method as described forcompound 153. Displacement of the brosylate was performed using6-methoxy-1-naphthol. The subsequent steps were analogous to previouslydescribed methods. The final product was purified by reverse phase HPLC(A: water/0.05% TFA, B: acetonitrile/0.5% TFA). ¹H NMR (300 MHz, CD₃OD):

8.13 (d, J=9 Hz, 1H), 7.40-7.35 (m, 2H), 7.30-7.16 (m, 6H), 7.01 (d,J=11 Hz, 1H), 6.85-6.80 (m, 1H), 5.73 (q, J=11 Hz, 1H), 5.42-5.34 (m,2H), 4.85-4.76 (m, 1H), 4.64-4.55 (m, 2H), 4.32 (d, J=8 Hz, 1H),4.06-3.97 (m, 1H), 3.90 (s, 3H), 3.39-3.11 (m, 2H), 2.87-2.70 (m, 1H),2.70-2.57 (m, 1H), 2.45-2.24 (m, 2H), 1.95-1.25 (m, 19H). ³¹P NMR (121.4MHz, CD₃OD):

43.13.

EI MS (m/z) 743.7 [M+H]⁺, 765.7 [M+Na]⁺.

Example 204 Preparation of Compound 204

Compound 204 was afforded as a white solid. ¹H NMR (300 MHz, CD₃OD):

8.33 (m, 1H), 7.90 (m, 1H), 7.55 (min, 1H), 7.³¹ (m, 3H), 6.94 (m, 2H),5.90 (bs, 1H), 5.73 (m, 1H), 5.33 (m, 1H), 4.74 (m, 2H), 4.42 (bs, 1H),4.13 (m, 2H), 4.00 (s, 3H), 3.57 (m, 1H), 2.76 (m, 2H), 2.57 (m, 1H)2.³¹ (m, 1H), 1.86 (m, 2H) 1.65-1.2 (m, 18H); ³¹P NMR (121.4 MHz,CD₃OD):

39.386

LC (6 minute run, r.t.=4.79 min) MS (781.2, M+1)

Example 205 Preparation of Compound 205

Compound 205 was afforded as a white solid. ¹H NMR (300 MHz, CD₃OD):

8.28 (d, J=9.2 Hz, 1H), 7.90 (d, J=6.4 Hz, 1H), 7.50 (d, J=6.8 Hz, 1H),7.³¹ (m, 3H), 6.98 (m, 2H), 5.89 (bs, 1H), 4.74 (m, 2H), 4.42 (bs, 1H),4.24 (m, 1H) 4.08 (m, 1H), 3.98 (s, 3H) 3.40 (m, 2H), 2.76 (m, 1H), 2.48(m, 1H) 1.99 (m, 1H), 1.79 (m, 2H) 1.65-1.2 (m, 22H); ³¹P NMR (121.4MHz, CD₃OD):

41.217. LC (6 minute run, r.t.=4.74 min) MS (783.2, M+1)

Example 206 Preparation of Compound 206

Compound 206 was prepared using a method similar to that described forcompound 185. LCMS (M+1): 953.32

Example 207 Preparation of Compound 207

Compound 207 was prepared using procedures similar to those describedherein. LCMS (M+1): 745.

Example 208 Preparation of Compound 208

Compound 208 was prepared using procedures similar to those describedherein. LCMS (M+1): 942.

Example 209 Preparation of Compound 209

a. Amino acid (702 mg, 3.16 mmol) was slurried in MeCN (31 mL) and satdNaHCO₃ (aq, 32 mL) was added followed byN-cyclopentyloxy-(Carbonyloxy)succinimide (1.00 g, 4.41 mmol),portion-wise. After 2 h at room temperature, the mixture was coned invacuo and partitioned between ethylacetate (40 mL)/H₂O (30 mL) whereuponit was acidified with HCl (1N) to pH=1. The aqueous layer was drawn offand extracted with ethylacetate (80 mL×2). The combined organic layerswere washed with brine (80 mL), dried over Na₂SO₄, and coned in vacuoaffording 1.32 g of (R)-3-methyl-8-nonenoic acid product contaminatedwith N-Cyclopentyloxy(Carbonyloxy)succinimide. LCMS (M+1): 297.88.

b. N-boc-4-cis-brosylproline methylester (1.77 g, 3.80 mmol) wasdissolved in DCM (16 mL) and HCl (4N in dioxane, 16 mL) was addedslowly. After 1.5 hours at room temperature the solution wasconcentrated in vacuo and the crude solid was dissolved in DMF (32 mL).To this solution was added crude (R)-3-methyl-8-nonenoic acid (1.32 g,3.16 mmol), and HATU (2.40 g, 6.32 mmol). The slurry was cooled to 0° C.and N-methylmorpholine (1.75 mL, 15.92 mmol) was added drop-wisewhereupon the cold bath was removed. After 16 hours at room temperature,the soln was poured into LiCl (2%, aq, 250 mL) and extracted withethylacetate (100 mL×3). The combined organic layers were washed withLiCl (2%, aq, 25 mL), satd NaHCO₃ (100 mL), satd NH₄Cl (100 mL), brine(100 mL), dried over MgSO₄, and coned in vacuo. The crude residue waspurified via silica-gel chromatography (ethylacetate-hexanes) to afford1.99 g of amide product in 98% yield. LCMS (M+1): 644.87.

c. To a soln of proline methylester (1.98 g, 3.07 mmol) in THF (10 mL)and MeOH (10 mL) was added a solution of LiOH (379 mg, 0.915 mmol, 10 mLH₂O) dropwise. After 2 hours at room temperature, the resultingsuspension was diluted with H₂O and acidified to pH=1. The aqueous wasextracted with ethylacetate (50 mL×4). The combined organic layers werewashed with brine (75 mL), dried over MgSO₄, and concentrated in vacuo.The crude solid was dissolved in DMF (30 mL) and aminophosphinate (1.13g, 3.74 mmol) was added followed by HATU (2.34 g, 6.14 mmol). The slurrywas cooled to 0° C. and N-methylmorpholine (1.70 mL, 15.46 mmol) wasadded drop-wise whereupon the cold bath was removed. After 16 hours atroom temperature, the solution was poured into LiCl (5%, aq, 200 mL) andextracted with ethylacetate (100 mL×3). The combined organic layers werewashed with LiCl (5%, aq, 100 mL), HCl (0.5 N, 100 mL), satd NaHCO₃ (100mL), brine (100 mL), dried over MgSO₄, and concentrated in vacuo. Thecrude residue was purified via silica-gel chromatography(ethylacetate-hexanes) to afford 2.17 g of amide product in 78% yield.LCMS (M+1): 912.02.

c. Phosphinate-diene was dissolved in CH₂Cl₂ (237 mL) and the solutionwas degassed for 30 minutes. The solution was heated to reflux andGrubb's G1 catalyst (492 mg, 0.598 mmol) was added. After 20 hours atreflux, trishydroxymethylphosphine (3.71 g, 29.90 mmol), TEA (8.6 mL,59.8 mmol), and H₂O (100 mL) were added and the reaction mixture wasrefluxed overnight. After cooling to room temperature, the layers wereseparated. The organic layer was washed with H₂O (100 mL), ½ satd NaHCO₃(100 mL×2), LiCl (5%, aq, 100 mL), brine (100 mL), dried over MgSO₄, andconcentrated in vacuo. The crude residue was purified by silica-gelchromatography (ethylacetate-hexanes) to afford 1.32 g of product in 63%yield. LCMS (M+1): 884.01.

d. Macrocycle (1.32 g, 1.49 mmol), hydroxyquinoline (523 mg, 1.49 mmol),and Cs₂CO₃ (973 mg, 2.98 mmol) were slurried in NMP (5.0 mL) and heatedto 65° C. for 8 hours. The mixture was poured into LiCl (5%, aq, 50 mL)and extracted with ethylacetate (30 mL×3). The combined organic layerswere washed with LiCl (5%, aq, mL×3), satd NaHCO₃ (40 mL), ½ satd NaHCO₃(50 mL), brine (50 mL), dried over Na₂SO₄, and coned in vacuo. The cruderesidue was purified via silica-gel chromatography(ethylacetate-hexanes) to afford 1.13 g of aminothiazole in 76% yield.LCMS (M+1): 997.31.

e. Aminothiazole (1.13 g, 1.13 mmol) was dissolved in MeCN (11.2 mL) andcooled to 0° C. (precipitation was observed). To the cooled slurry wasadded trimethylsilyliodide (800 μL, 5.62 mmol) drop-wise, whereupon thecold-bath was removed. After 35 minutes at room temperature, thesolution was cooled to 0° C. and 2,6-lutidine (1.3 mL, 11.2 mmol) wasadded followed by MeOH (1.3 mL). The soln was concentrated in vacuo andpurified via reverse-phase HPLC (MeCN—H₂O with 0.1% TFA) affording 1.002g of phosphinic acid in 80% yield. ¹H NMR (300 MHz, CD₃OD) d 8.33 (d,J=9.9 Hz, 1H), 8.32 (s, 1H), 7.87 (s, 1H), 7.65 (d, J=9.3 Hz, 1H),7.33-7.21 (m, 1H), 6.94 (dd, J=7.8, 7.5 Hz, 2H), 5.85 (s, 1H), 5.76-5.66(m, 1H), 5.35 (dd, J=9.9 Hz, 1H), 4.73 (dd, J=9.3, 7.5 Hz, 1H), 4.18 (s,3H), 4.15-4.01 (m, 3H), 3.83-3.77 (m, 1H), 3.64 (s, 3H), 3.62-3.55 (m,1H), 2.92-2.81 (m, 1H), 2.78-2.58 (m, 2H), 2.48-2.32 (m, 1H), 2.07-1.91(m, 1H), 1.84-1.76 (m, 1H), 1.75-1.22 (m, 23H), 1.19-1.01 (m, 2H),0.97-0.90 (m, 3H); (M+1): 969.42.

Example 210 Preparation of Compound 210

Phosphinic acid (702 mg, 0.632 mmol), sodium acetate (778 mg, 9.48mmol), and tosylhydrazide (886 mg, 4.76 mmol), were slurried in DME(5.75 mL) and H₂O (575 μL) and heated to 95° C. for 1.5 hours. Afterwhich, more sodium acetate (160 mg, 1.95 mmol) and tosylhydrazide (177mg, 0.95 mmol) were added and the mixture was heated at 95° C. foranother 1.2 hours. It was coned in vacuo, dissolved in MeOH, filtered,and purified via reverse-phase HPLC (MeCN—H₂O with 0.1% TFA) affording371 mg of phosphinic acid in 53% yield. ¹H NMR (300 MHz, CD₃OD)

8.32 (t, J=4.5 Hz, 2H), 7.88 (s, 1H), 7.66 (d, J=9.3 Hz, 1H), 7.38-7.25(m, 1H), 6.98 (dd, J=7.8 Hz, 2H), 5.85 (s, 1H), 4.73 (dd, J=8.7, 8.4 Hz,2H), 4.17 (s, 3H), 4.12-3.98 (m, 3H), 3.79 (d, J=10.5 Hz, 1H), 3.73-3.67(m, 1H), 3.57-3.50 (m, 1H), 3.43 (d, J=10.5 Hz, 1H), 2.94-2.85 (m, 1H),2.58-2.47 (m, 1H), 2.04-1.91 (m, 1H), 1.85-1.04 (m, 24H) 1.37 (d, J=6.6Hz, 6H), 0.90 (d, J=6.6 Hz, 3H); LCMS (M+1): 971.28.

Example 211 Preparation of Compound 211

a. Proline methylester (4.85 g, 8.67 mmol) was dissolved in MeOH (30 mL)and THF (30 mL). To the organic soln was slowly added a solution oflithium hydroxide (3.60 g, 86.7 mmol) in H₂O (30 mL). The mixture wasstirred at room temperature for 1 hour, whereupon it was diluted withH₂O (20 mL), acidified with HCl (1N) to pH=2, and extracted withethylacetate (40 mL×3). The combined organic layers were dried overMgSO₄ and coned in vacuo affording 4.55 g of acid in 96% crude yield.

b. Vinylcyclopropane phosphinate (5.90 g, 13.6 mmol) was dissolved inTHF (100 mL) and borane-THF complex (1.0 M in THF, 13.6 mL, 13.6 mmol)was slowly added. After 2 hours at room temperature, hydrogen peroxide(30% in water, 1.4 mL, 13.6 mmol) was slowly added followed by sodiumhydroxide (1.0 M, 17.7 mL, 17.7 mmol) and the mixture was stirred for anadditional 1 hour at room temperature. The mixture was then diluted withH₂O (100 ml) and extracted with ethylacetate (100 mL×3). The combinedorganic layers were washed with brine (100 mL), dried over Na₂SO₄, andconed in vacuo. The crude residue was purified via silica-gelchromatography (ethylacetate-hexanes) to afford 4.19 g of alcohol in 68%yield.

c. Amino alcohol (1.57 g, 3.47 mmol), allylbromide (1.50 mL, 17.3 mmol),and mol sieves (4 Å) were stirred in DCM (17 mL) for 30 min. To themixture was added silver oxide (2.80 g, 12.14 mmol) and it was stirredat room temperature overnight. The mixture was filtered and concentratedin vacuo. The crude residue was purified via silica-gel chromatography(ethylacetate-hexanes) to afford 465 mg of allylated product in 27%yield.

d. Aminoether (1.43 g, 2.89 mmol) was stirred in TFA (6 mL) anddimethylsulfide (2 mL) at room temperature overnight. The solution wasdiluted with isopropylacetate (30 mL)/heptanes (30 mL) and extractedwith HCl (1N, 30 mL×2). The organic layer was further diluted withheptanes (30 mL) and extracted with HCl (1 N, 30 mL). The dilution withheptanes and extraction with HCl was repeated 2×. The combined aqueouslayers were basified with sodium hydroxide to pH=12 and extracted withethyl acetate (100 mL×3). The combined organic layers were washed withbrine (100 mL), dried over Na₂SO₄, and concentrated in vacuo to afford754 mg of amine in 72% yield. LCMS (M+1): 360.07.

e. Proline acid (303 mg, 0.555 mmol) and TEA (85 μL, 0.61 mmol) weredissolved in THF (4.0 mL) and cooled to 0° C. To the solution was addedisobutylchloroformate (80 μL, 0.61 mmol). After an additional 40 minutesat 0° C., the amine (200 mg, 0.555 mmol) was added as a solution in THF(1.5 mL) and the mixture was allowed to warm to room temperature.Following 2 hours at room temperature, the mixture was diluted with satdNaHCO₃ and extracted with ethylacetate. The combined organic layers werewashed with brine, dried over MgSO₄ and coned in vacuo. The cruderesidue was purified via silica-gel chromatography(ethylacetate-hexanes) to afford 306 mg of amide product in 62% yield.LCMS (M+1): 889.87.

f. Phosphinate-diene (986 mg, 1.11 mmol) was dissolved in DCM (100 mL)and the solution was degassed for 30 minutes. The solution was heated toreflux and Grubb's G1 catalyst (250 mg, 0.31 mmol) was added. After 16hours at reflux, more Grubb's G1 catalyst (45 mg, 0.055 mmol) was added.After an additional 3 hours at reflux more Grubb's G1 catalyst (45 mg,0.055 mmol) was added. After an additional 3 hours at refluxtrishydroxymethylphosphine (2.3 g, 18.5 mmol), TEA (5.1 mL, 37 mmol),and H₂O (20 mL) were added and the reaction mixture was refluxedovernight. After cooling to room temperature, the layers were separated.The aqueous layer was washed with DCM (50 mL), dried over Na₂SO₄, andconcentrated in vacuo. The crude residue was purified via silica-gelchromatography (ethylacetate-hexanes) to afford 440 mg of product in 46%yield. LCMS (M+1): 859.93.

g. Macrocycle (532 mg, 0.62 mmol), hydroxyquinoline (216 mg, 0.62 mmol),and Cs₂CO₃ (404 mg, 1.24 mmol) were slurried in NMP (6.0 mL) and heatedto 65° C. for 8 hours. The mixture was poured into LiCl (5%, aq, 60 mL)and extracted with ethylacetate (30 mL×3). The combined organic layerswere washed with LiCl (5%, aq, 35 mL×3), satd NaHCO₃ (40 mL), brine (50mL), dried over Na₂SO₄, and coned in vacuo. The crude residue waspurified via silica-gel chromatography (methanol-ethylacetate) to afford456 mg of aminothiazole in 76% yield. LCMS (M+1): 973.27.

h. Macrocycle (456 mg, 0.468 mmol) was dissolved in MeCN (5.0 mL) andTMSI (0.34 mL, 2.35 mmol) was added drop-wise. After 10 minutes at roomtemperature, 2,6-lutidine (0.27 mL, 2.33 mmol) was added followed byMeOH (0.27 mL) and the solution was coned in vacuo. The crude residuewas dissolved in MeCN (3.0 mL) and satd NaHCO₃ (3.0 mL) was added. Tothe bilayer was added N-Cyclopentyloxy(Carbonyloxy) succinimide (127 mg,0.56 mmol). After 2 hours at rt the layers were separated and theaqueous layer was acidified to pH=2 and extracted with ethylacetate (7mL×3). The combined organic layers were coned in vacuo and the cruderesidue was dissolved in DMF and purified directly by reverse-phasepreparative HPLC (Column: Phenomenex Gemini 5u, C18, 110A, 75×30 mm,Gradient: 30-95% acetonitrile-water with 0.1% TFA) affording 216 mg ofphosphinic acid in 48% yield. LCMS (M+1): 957.20.

Example 212 Preparation of Compound 212

Phosphinic acid (210 mg, 0.22 mmol), sodium acetate (270 mg, 3.29 mmol),and tosylhydrazide (310 mg, 1.65 mmol), were slurried in DME (5.0 mL)and H₂O (500 μL) and heated to 95° C. for 2 hours. The mixture wascooled to 0° C. and HCl (6N, 550 μL, 3.29 mmol) was added and it wasconcentrated in vacuo, dissolved in MeOH, filtered, and purified viareverse-phase HPLC (Column: Phenomenex Gemini 5u, C18, 110A, 75×30 mm,Gradient: 30-95% acetonitrile-water with 0.1% TFA) affording 170 mg ofphosphinic acid in 80% yield. LCMS (M+1): 959.33.

Example 213 Preparation of Compound 213

To a solution of (A) (250 mg, 0.272 mmol) in CH₃CN (3 mL, 0.1 M) at 0°C. was added iodotrimethylsilane (194 μL, 1.36 mmol). The reactionmixture was stirred at 0° C. for 5 minutes. 2,6-Lutidine (315 μL, 2.72mmol) was added and stirred for 1.5 hours. MeOH was added and stirredfor 30 minutes. The mixture was concentrated and re-dissolved in minimalMeOH and purified by reverse phase HPLC to afford 190 mgs (70%) of (B)as a TFA salt. ¹H NMR (300 MHz, CDCl₃):

15-8.0 (m, 4H), 7.49 (d, J=7.3, 3H), 7.26-7.04 (m, 5H), 6.74 (dd, J=7.1Hz, J=8.2 Hz, 2H), 5.68 (d, J=8.2 Hz, 1H), 5.48 (m, 1H), 4.85 (m, 1H),4.62-4.36 (m, 3H), 4.25-3.99 (m, 5H), 3.48 (t, J=15.3 Hz, 1H), 3.21 (t,J=16.5 Hz, 1H), 2.60 (m, 1H), 2.16 (m, 1H), 1.73-1.23 (m, 22H); ³¹P (75MHz, CDCl₃):

42.04. LC/MS: M+1=891.

Example 213 Preparation of Compound 213

To a solution of (C) (35 mg, 0.039 mmol) in DME (1 mL)/H₂O (0.1 mL) wasadded p-tosylhydrazide (37 mg, 0.196 mmol) and NaOAc (32 mg, 0.39 mmol).The reaction mixture was heated to 95° C. for 1.5 hour and cooled toroom temperature. A few drops of 1 N HCl was added to adjust the pH=2.The crude product was purified by reverse phase HPLC to give 12.4 mg ofacid (D) in 36% yield. ¹H NMR (300 MHz, CDCl₃):

8.32-8.24 (m, 2H), 7.92 (d, J=6.4 Hz, 2H), 7.56-7.27 (m, 5H), 7.00 (m,1H), 6.71-6.66 (m, 2H), 5.73 (s, 1H), 5.54 (d, J=7.3 Hz, 1H), 4.76-4.73(m, 2H), 4.47-4.43 (m, 2H), 4.14 (s, 3H), 3.39 (t, J=15.6 Hz, 1H), 3.24(t, J=14.6 Hz, 1H), 2.61 (m, 1H), 1.79-1.19 (m, 3H); ³¹P (75 MHz, CDCl₃)d 42.77 (s, 1H). LC/MS: M+1=893.

Example 214 Preparation of Compounds 214-225

Using procedures similar to those described herein, the followingcompounds 214-225 can also be prepared.

Biological Assays

NS3 Enzymatic Potency:

Purified NS3 protease is complexed with NS4A peptide and then incubatedwith serial dilutions of compound (DMSO used as solvent). Reactions arestarted by addition of dual-labeled peptide substrate and the resultingkinetic increase in fluorescence is measured. Non-linear regression ofvelocity data is performed to calculate IC₅₀s. Activity is initiallytested against genotype 1b protease. Depending on the potency obtainedagainst genotype 1b, additional genotypes (1a, 2a, 3) and/or proteaseinhibitor resistant enzymes (D168Y, D 68V, or A156T mutants) may betested. BILN-2061 is used as a control during all assays. Representativecompounds of the invention were evaluated in this assay and weretypically found to have IC₅₀ values of less than about 1 μm.

Replicon Potency and Cytotoxicity:

Huh-luc cells (stably replicating Bartenschlager's1389luc-ubi-neo/NS3-3′/ET genotype 1b replicon) are treated with serialdilutions of compound (DMSO is used as solvent) for 72 hours. Repliconcopy number is measured by bioluminescence and non-linear regression isperformed to calculate EC₅₀s. Parallel plates treated with the same drugdilutions are assayed for cytotoxicity using the Promega CellTiter-Glocell viability assay. Depending on the potency achieved against the 1breplicon, compounds may be tested against a genotype 1a replicon and/orinhibitor resistant replicons encoding D168Y or A156T mutations.BILN-2061 is used as a control during all assays. Representativecompounds of the invention were evaluated in this assay and weretypically found to have EC₅₀ values of less than about 5 μm.

Effect of Serum Proteins on Replicon Potency

Replicon assays are conducted in normal cell culture medium (DMEM+10%FBS) supplemented with physiologic concentrations of human serum albumin(40 mg/mL) or a-acid glycoprotein (1 mg/mL). EC₅₀s in the presence ofhuman serum proteins are compared to the EC₅₀ in normal medium todetermine the fold shift in potency.

Enyzmatic Selectivity:

The inhibition of mammalian proteases including Porcine PancreaticElastase, Human Leukocyte Elastase, Protease 3, and Cathepsin D aremeasured at K_(m) for the respective substrates for each enzyme. IC₅₀for each enzyme is compared to the IC₅₀ obtained with NS3 1b protease tocalculate selectivity. Representative compounds of the invention haveshown activity.

MT-4 Cell Cytotoxicity:

MT4 cells are treated with serial dilutions of compounds for a five dayperiod. Cell viability is measured at the end of the treatment periodusing the Promega CellTiter-Glo assay and non-linear regression isperformed to calculate CC₅₀.

Compound Concentration Associated with Cells at EC₅₀:

Huh-luc cultures are incubated with compound at concentrations equal toEC₅₀. At multiple time points (0-72 hours), cells are washed 2× withcold medium and extracted with 85% acetonitrile; a sample of the mediaat each time-point will also be extracted. Cell and media extracts areanalyzed by LC/MS/MS to determine the Molar concentration of compoundsin each fraction. Representative compounds of the invention have shownactivity.

Solubility and Stability:

Solubility is determined by taking an aliquot of 10 mM DMSO stocksolution and preparing the compound at a final concentration of 100 μMin the test media solutions (PBS, pH 7.4 and 0.1 N HCl, pH 1.5) with atotal DMSO concentration of 1%. The test media solutions are incubatedat room temperature with shaking for 1 hr. The solutions will then becentrifuged and the recovered supernatants are assayed on the HPLC/UV.Solubility will be calculated by comparing the amount of compounddetected in the defined test solution compared to the amount detected inDMSO at the same concentration. Stability of compounds after a 1 hourincubation with PBS at 37° C. will also be determined.

Stability in Cryopreserved Human, Dog, and Rat Hepatocytes:

Each compound is incubated for up to 1 hour in hepatocyte suspensions(100 μl, 80,000 cells per well) at 37° C. Cryopreserved hepatocytes arereconstituted in the serum-free incubation medium. The suspension istransferred into 96-well plates (50 μL/well). The compounds are dilutedto 2 μM in incubation medium and then are added to hepatocytesuspensions to start the incubation. Samples are taken at 0, 10, 30 and60 minutes after the start of incubation and reaction will be quenchedwith a mixture consisting of 0.3% formic acid in 90% acetonitrile/10%water. The concentration of the compound in each sample is analyzedusing LC/MS/MS. The disappearance half-life of the compound inhepatocyte suspension is determined by fitting the concentration-timedata with a monophasic exponential equation. The data will also bescaled up to represent intrinsic hepatic clearance and/or total hepaticclearance.

Stability in Hepatic S9 Fraction from Human, Dog, and Rat:

Each compound is incubated for up to 1 hour in S9 suspension (500 μl, 3mg protein/mL) at 37° C. (n=3). The compounds are added to the 59suspension to start the incubation. Samples are taken at 0, 10, 30, and60 minutes after the start of incubation. The concentration of thecompound in each sample is analyzed using LC/MS/MS. The disappearancehalf-life of the compound in S9 suspension is determined by fitting theconcentration-time data with a monophasic exponential equation.

Caco-2 Permeability:

Compounds are assayed via a contract service (Absorption Systems, Exton,Pa.). Compounds are provided to the contractor in a blinded manner. Bothforward (A-to-B) and reverse (B-to-A) permeability will be measured.Caco-2 monolayers are grown to confluence on collagen-coated,microporous, polycarbonate membranes in 12-well Costar Transwell®plates. The compounds are dosed on the apical side for forwardpermeability (A-to-B), and are dosed on the basolateral side for reversepermeability (B-to-A). The cells are incubated at 37° C. with 5% CO₂ ina humidified incubator. At the beginning of incubation and at 1 hr and 2hr after incubation, a 200-μL aliquot is taken from the receiver chamberand replaced with fresh assay buffer. The concentration of the compoundin each sample is determined with LC/MS/MS. The apparent permeability,Papp, is calculated.

Plasma Protein Binding:

Plasma protein binding is measured by equilibrium dialysis. Eachcompound is spiked into blank plasma at a final concentration of 2 μM.The spiked plasma and phosphate buffer is placed into opposite sides ofthe assembled dialysis cells, which will then be rotated slowly in a 37°C. water bath. At the end of the incubation, the concentration of thecompound in plasma and phosphate buffer is determined. The percentunbound is calculated using the following equation:

${\%\mspace{14mu}{Unbound}} = {100 \cdot ( \frac{C_{f}}{C_{b} + C_{f}} )}$Where C_(f) and C_(b) are free and bound concentrations determined asthe post-dialysis buffer and plasma concentrations, respectivelyCYP450 Profiling:

Each compound is incubated with each of 5 recombinant human CYP450enzymes, including CYP1A2, CYP2C9, CYP3A4, CYP2D6 and CYP2C19 in thepresence and absence of NADPH. Serial samples will be taken from theincubation mixture at the beginning of the incubation and at 5, 15, 30,45 and 60 min after the start of the incubation. The concentration ofthe compound in the incubation mixture is determined by LC/MS/MS. Thepercentage of the compound remaining after incubation at each time pointis calculated by comparing with the sampling at the start of incubation.

Stability in Rat, Dog, Monkey and Human Plasma:

Compounds will be incubated for up to 2 hour in plasma (rat, dog,monkey, or human) at 37° C. Compounds are added to the plasma at finalconcentrations of 1 and 10 ug/mL. Aliquots are taken at 0, 5, 15, 30,60, and 120 min after adding the compound. Concentration of compoundsand major metabolites at each timepoint are measured by LC/MS/MS.

All publications, patents, and patent documents are incorporated byreference herein, as though individually incorporated by reference. Theinvention has been described with reference to various specific andpreferred embodiments and techniques. However, it should be understoodthat many variations and modifications may be made while remainingwithin the spirit and scope of the invention.

What is claimed:
 1. A compound of formula I:

or a pharmaceutically acceptable salt thereof, wherein: R² is selectedfrom, a) —C(Y¹)(A³), b) (C2-10)alkyl, (C3-7)cycloalkyl or(C1-4)alkyl-(C3-7)cycloalkyl, where said cycloalkyl and alkyl-cycloalkylmay be optionally mono-, di- or tri-substituted with (C1-3)alkyl, orwhere said alkyl, cycloalkyl and alkyl-cycloalkyl may optionally bemono- or di-substituted with substituents selected from hydroxy andO—(C1-4)alkyl, or where each of said alkyl-groups may optionally bemono-, di- or tri-substituted with halogen, or where each of saidcycloalkyl groups being 5-, 6- or 7-membered, one or two —CH₂-groups notbeing directly linked to each other may be optionally substitutedreplaced by —O— such that the O-atom is linked to the N atom to which R²is attached via at least two C-atoms, c) phenyl, (C1-3)alkyl-phenyl,heteroaryl or (C1-3)alkyl-heteroaryl, wherein the heteroaryl-groups are5- or 6-membered having from 1 to 3 heteroatoms selected from N, O andS, wherein said phenyl and heteroaryl groups may optionally be mono-,di- or trisubstituted with substituents selected from halogen, —OH,(C1-4)alkyl, O—(C1-4)alkyl, S—(C1-4)alkyl, —NH₂, —CF₃, —NH((C1-4)alkyl)and —N((C1-4)alkyl)₂, —CONH₂ and —CONH—(C1-4)alkyl; and wherein said(C1-3)alkyl may optionally be substituted with one or more halogen; ord) —S(O)₂(A³); R³ is H or (C1-6)alkyl; Y¹ is independently O or S; Z isO, S, or NR³; Z¹ is:

in which: R_(a) is (C1-6)alkoxy; R_(b) is H, or Cl; R_(c) is phenyl thatis optionally substituted with one or more F, Cl, Br, I, (C1-6)alkyl, or(C1-6)alkoxy; R_(d) and R_(e) are each independently H or (C1-6)alkyl;Q¹ and Z^(2a) taken together with the atoms to which they are attachedform heterocycle, which heterocycle may optionally be substituted withone or more oxo (═O); Z^(2b) is H, (C1-6)alkyl, (C2-8)alkenyl,(C2-8)alkynyl; A³ is independently selected from H, —OH, —C(O)OH, cyano,alkyl, alkenyl, alkynyl, amino, amido, imido, imino, halogen, CF₃,CH₂CF₃, cycloalkyl, nitro, aryl, aralkyl, alkoxy, aryloxy, heterocycle,—C(A²)₃, —C(A²)₂-C(O)A², —C(O)A², —C(O)OA², —O(A²), —N(A²)₂, —S(A²),—CH₂P(Y¹)(A²)(OA²), —CH₂P(Y¹)(A²)(N(A²)₂), —CH₂P(Y¹)(OA²)(OA²),—OCH₂P(Y¹)(OA²)(OA²), —OCH₂P(Y¹)(A²)(OA²), —OCH₂P(Y¹)(A²)(N(A²)₂),—C(O)OCH₂P(Y¹)(OA²)(OA²), —C(O)OCH₂P(Y¹)(A²)(OA²),—C(O)OCH₂P(Y¹)(A²)(N(A²)₂), —CH₂P(Y¹)(OA²)(N(A²)₂),—OCH₂P(Y¹)(OA²)(N(A²)₂), —C(O)OCH₂P(Y¹)(OA²)(N(A²)₂),—CH₂P(Y¹)(N(A²)₂)(N(A²)₂), —C(O)OCH₂P(Y¹)N(A²)₂)(N(A²)₂),—OCH₂P(Y¹)(N(A²)₂)(N(A²)₂), —(CH₂)_(m)-heterocycle,—(CH₂)_(m)C(O)Oalkyl, —O—(CH₂)_(m)—O—C(O)—Oalkyl,—O—(CH₂)_(r)O—C(O)—(CH₂)_(m)-alkyl, —(CH₂)_(m)O—C(O)—O-alkyl,—(CH₂)_(m)O—C(O)—O-cycloalkyl, —N(H)C(Me)C(O)O-alkyl, or alkoxyarylsulfonamide, wherein each A³ is optionally substituted with 1 to 4—P(Y¹)(OA²)(OA²), —P(Y¹)(OA²)(N(A²)₂), —P(Y¹)(A²)(OA²),—P(Y¹)(A²)(N(A²)₂), or P(Y¹)(N(A²)₂)(N(A²)₂), —C(═O)N(A²)₂), halogen,alkyl, alkenyl, alkynyl, aryl, carbocycle, heterocycle, aralkyl, arylsulfonamide, aryl alkylsulfonamide, aryloxy sulfonamide, aryloxyalkylsulfonamide, aryloxy arylsulfonamide, alkyl sulfonamide, alkyloxysulfonamide, alkyloxy alkylsulfonamide, arylthio, —(CH₂)_(m)heterocycle,—(CH₂)_(m)—C(O)O-alkyl, —O(CH₂)_(m)OC(O)Oalkyl,—O—(CH₂)_(m)—O—C(O)—(CH₂)_(m)-alkyl, —(CH₂)_(m)—O—C(O)—O-alkyl,—(CH₂)_(m)—O—C(O)—O-cycloalkyl, —N(H)C(CH₃)C(O)O-alkyl, or alkoxyarylsulfonamide; A² is independently selected from H, alkyl, alkenyl,alkynyl, amino, amino acid, alkoxy, aryloxy, cyano, haloalkyl,cycloalkyl, aryl, heteroaryl, alkylsulfonamide, or arylsulfonamide; andm is 0 to
 6. 2. The compound of claim 1, wherein R² is —C(O)(OA²) and Zand Y¹ are O.
 3. The compound of claim 2, wherein R_(c) is phenyloptionally substituted with halo, alkyl of 1-6 carbon atoms, or alkoxyof 1-6 carbon atoms.
 4. The compound of claim 3, wherein A² iscycloalkyl of 3-8 carbon atoms and A³ is hydrogen or alkyl of 1-6 carbonatoms optionally substituted with phenyl or a heterocycle.
 5. Thecompound of claim 4, wherein Z^(2b) is hydrogen.
 6. A compound selectedfrom the group consisting of:

or a pharmaceutically acceptable salt thereof.